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title="Jump to previuos match">&#9664;</a><button id="jr-fip-matches">no matches yet</button><a id="jr-fip-next" class="wsprkl btn" title="Jump to next match">&#9654;</a></nav></nav></div><div id="jr-epub-interstitial" class="hidden"></div><div id="jr-content"><article data-type="main"><div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><div class="fm-sec"><h1 id="_NBK20266_"><span class="label">Chapter 7</span><span class="title" itemprop="name">Evolution of Central Metabolic Pathways: The Playground of Non-Orthologous Gene Displacement</span></h1><p class="fm-aai"><a href="#_NBK20266_pubdet_">Publication Details</a></p></div></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><p>One of the central goals of functional genomics is the complete reconstruction of the
metabolic pathways of the organisms, for which genome sequences have been obtained. As
discussed in <a href="/books/n/sef/A4/?report=reader">Chapter 1</a>, there is no chance
that all necessary biochemical experiments are ever done in any substantial number of
organisms. Therefore, reconstructions made through comparative genomics, combined with
the knowledge derived from experiments on model systems, are the only realistic path to
a satisfactory understanding of the biochemical diversity of life and to the
characterization of poorly studied and hard-to-grow organisms (including extremely
important ones, e.g. the syphilis spirochete <i>T. pallidum</i> [<a href="/books/n/sef/A727/?report=reader#A971">243</a>,<a href="/books/n/sef/A727/?report=reader#A1615">887</a>]).</p><p>In the pre-genomic era, metabolic reconstruction might have seemed to be a relatively
easy task, given the overall similarity of the key metabolic enzymes in several model
organisms, such as <i>E. coli</i>, <i>B. subtilis</i>, yeast,
plants, and animals. Although cases of non-orthologous (unrelated or distantly related)
enzymes catalyzing the same reaction, such as the two distinct forms of
fructose-1,6-bisphosphate aldolases, phosphoglycerate mutases, and superoxide
dismutases, have been known for a long time, these cases were generally perceived as
rare and, more or less, inconsequential [<a href="/books/n/sef/A727/?report=reader#A915">187</a>,<a href="/books/n/sef/A727/?report=reader#A986">258</a>,<a href="/books/n/sef/A727/?report=reader#A999">271</a>,<a href="/books/n/sef/A727/?report=reader#A1277">549</a>]. The
availability of complete genomes is gradually changing this perception, making us
realize just how common these cases of analogous (as opposed to homologous) enzymes are
in nature (see <a href="/books/n/sef/A22/?report=reader#A40">2.2.5</a>). The phenomenon of
non-orthologous gene displacement turned out to be a major complication (but also a
major source of unexpected findings) for the analysis of metabolic pathways, making it
particularly hard to automate. Indeed, whenever an ortholog of a given metabolic enzyme
from the model organisms is not detected in the organism of interest (the initial step
of metabolic reconstruction, the identification of orthologs of known enzymes, can be
automated almost completely), the process turns into &#x0201c;detective
work&#x0201d;. The researcher needs to identify a set of gene products that, on the
basis of their predicted biochemical activities, potentially could catalyze the reaction
in question. Often, there is more than one such candidate, and the choice between these
might not be possible without direct experiments. Furthermore, there is always a chance
that, however plausible, all candidates detected in such searches are false, whereas the
true culprit is a complete unknown. This makes metabolic reconstruction in the era of
comparative genomics a less precise but much more exciting undertaking.</p><p>In this chapter, we show how a COG-based reconstruction of bacterial and archaeal
metabolism helps organizing the existing data on microbial biochemistry, illuminates the
remaining questions, suggests candidates for some of the &#x0201c;missing&#x0201d;
enzymatic activities, and predicts the existence of novel enzymes that remain to be
discovered. For each metabolic reaction, we list the COGs that are known to catalyze it
or can be reasonably predicted to do so. We then compare the phyletic patterns of the
corresponding COGs to see if the current set of COGs is sufficient to suggest candidate
proteins to catalyze the given reaction in each organism with sequenced genome or still
unexplained gaps remain in metabolic pathways.</p><div id="A372"><h2 id="_A372_">7.1. Carbohydrate Metabolism</h2><div id="A373"><h3>7.1.1. Glycolysis</h3><p>We have already used the COG approach to demonstrate the complementarity of the
phyletic patterns of the three forms of phosphoglycerate mutase (see <a href="/books/n/sef/A22/?report=reader#A43">2.2.6</a>). <a href="/books/n/sef/A517/?report=reader#A520">Figure 7.1</a> shows the COGs that are known or predicted to include
glycolytic enzymes and shows their phyletic patterns. This superposition of COGs
and metabolic pathways provides a convenient framework for a detailed analysis
of the phylogenetic distribution of each of the glycolytic enzymes and the
general principles of evolution of carbohydrate metabolism. This figure shows,
for example, that <i>R. prowazekii</i>, an obligate intracellular
parasite and a relative of the mitochondria [<a href="/books/n/sef/A727/?report=reader#A758">30</a>], does not encode a single glycolytic enzyme. In contrast, all
other organisms with completely sequenced genomes encode enzymes of the lower
(tri-carbon) part of the pathway. This supports the notion that glycolysis is
the central pathway of carbohydrate metabolism and makes comparative analysis of
variants of this pathway all the more interesting.</p><div id="A374"><h4>Glucokinase (EC 2.7.1.2 )</h4><p>Fermentation of glucose starts with its phosphorylation, which is catalyzed
by glucokinase. Although many bacteria bypass the glucokinase step by
phosphorylating glucose concomitantly with its uptake by the PEP-dependent
phosphotransferase system, some of them, including <i>E. coli</i>,
encode a glucokinase (COG0837) that shares little sequence similarity with
yeast and human enzymes. There is also another bacterial form, found in
<i>S. coelicolor</i>, <i>Bacillus megaterium</i>,
and other bacteria [<a href="/books/n/sef/A727/?report=reader#A760">32</a>,<a href="/books/n/sef/A727/?report=reader#A1523">795</a>].</p><p>Recently, <i>P. furiosus</i> has been reported to encode an
ADP-dependent glucokinase [<a href="/books/n/sef/A727/?report=reader#A1163">435</a>].
This enzyme has no detectable sequence similarity to any other glucokinase
but shows significant structural similarity to enzymes of the ribokinase
family [<a href="/books/n/sef/A727/?report=reader#A1111">383</a>]. In retrospect,
several conserved motifs were detected in this new glucokinase and the
ribokinase family proteins, which indicates a homologous relationship. Thus,
a clear-cut case of non-orthologous gene displacement is observed: a
ribokinase family enzyme has been recruited to replace the typical
glucokinase. So far, the ADP-dependent glucokinase has been found only in
<i>M. jannaschii</i> and in pyrococci. The existence of at
least three distinct forms of glucokinase is remarkable, especially given
that this is apparently not an essential component of glycolysis. Moving
down the glycolytic pathway, we find similar examples of non-orthologous
gene displacement for several other, essential enzymes.</p></div><div id="A375"><h4>Glucose-6-phosphate isomerase (EC 5.3.1.9)</h4><p>Bacteria and eukaryotes encode several distinct but homologous forms of
glucose-6-phosphate isomerase (phosphoglucomutase) [<a href="/books/n/sef/A727/?report=reader#A1352">624</a>]. The classical (<i>E. coli</i>) form
of the enzyme is found in Gram-negative bacteria and in the cytoplasm of the
eukaryotic cell. A divergent version of this enzyme is found in
Gram-positive bacteria including <i>B. subtilis</i>, in <i>T.
maritima</i>, and some archaea, such as <i>M.
jannaschii</i> and <i>Halobacterium</i> sp. [<a href="/books/n/sef/A727/?report=reader#A1194">466</a>,<a href="/books/n/sef/A727/?report=reader#A1489">761</a>]. The most divergent members of this family of
glucose-6-phosphate isomerases were detected in <i>A. aeolicus</i>
and another subset of archaea, including <i>M.
thermoautotrophicum</i>, <i>A. pernix</i>, and
<i>Thermoplasma</i> spp. No enzyme of this family seems to be
encoded in the genomes of <i>A. fulgidus</i> or pyrococci.
Instead, <i>P. furiosus</i> has been shown to encode a novel
glucose-6-phosphate isomerase, which has highly conserved orthologs in
<i>P. horikoshii</i> and in <i>A. fulgidus</i>, but
so far not in any other organism [<a href="/books/n/sef/A727/?report=reader#A1059">331</a>]. Thus, two non-orthologous (in fact, apparently unrelated)
versions of this enzyme together account for the phosphoglucomutase activity
in all known microbial genomes, with the exception of <i>R.
prowazekii</i> and <i>U. urealyticum</i>. As indicated
above, the former does not encode any glycolytic enzymes, whereas the latter
apparently obtains fructose-6-phosphate by importing fructose concomitantly
with its phosphorylation through the fructose-specific phosphotransferase
system, thus bypassing the phosphoglucomutase stage.</p></div><div id="A376"><h4>Phosphofructokinase (EC 2.7.1.11)</h4><p>The next glycolytic enzyme, phosphofructokinase, offers an even more
interesting example of non-orthologous gene displacement. It is also an
example of an enzyme where several &#x0201c;missing&#x0201d; enzyme
forms have been discovered just in the past year.</p><p>The most common version of this enzyme, PfkA, is an ATP-dependent kinase of
unique structure found in bacteria and many eukaryotes. Plants have a
homologous enzyme, which, however, uses pyrophosphate as the phosphate
donor. Altogether, homologs of PfkA are found in nearly all bacteria and
eukaryotes but are conspicuously missing in <i>H. pylori</i> and
in all archaeal genomes sequenced so far. In addition, <i>E.
coli</i> encodes a second phosphofructokinase, PfkB, which is
unrelated to PfkA and instead belongs to the ribokinase family of
carbohydrate kinases.</p><p>A unique ADP-dependent phosphofructokinase has been described in <i>P.
furiosus</i> [<a href="/books/n/sef/A727/?report=reader#A1581">853</a>].
However, this enzyme appears to have a limited phyletic distribution: so
far, it was found only in <i>M. jannaschii</i> and in pyrococci.
This left the phosphofructokinase activity in other archaea unaccounted for
and suggested that additional forms of this enzyme might exist. Very
recently, a new ATP-dependent phosphofructokinase, which is a member of the
ribokinase family but is not specifically related to PfkB, has been
identified in <i>A. pernix</i> [<a href="/books/n/sef/A727/?report=reader#A1440">712</a>]. Close homologs of this protein (APE0012) were found in
<i>Halobacterium</i> sp., <i>A. fulgidus</i>,
<i>M. thermoautotrophicum</i>, and several other archaea.
Therefore, it seems likely that these ribokinase family enzymes function as
phosphofructokinases in all these archaea. Finally,
<i>Thermoplasma</i> does not encode orthologs of any of the
four forms of phosphofructokinase described above. This leaves two
possibilities: either thermoplasmas lack phosphofructokinase altogether
(along with fructose-1,6-bisphosphate aldolase; see below), or they might
have yet another, fifth variant of this enzyme.</p></div><div id="A377"><h4>Fructose-1,6-bisphosphate aldolase (EC 4.1.2.13)</h4><p>For more than 50 years now, it has been known that fructose-1,6-bisphosphate
aldolase exists in two distinct forms, a metal-independent one (class I) in
multicellular eukaryotes and a metal-dependent one (class II) in bacteria
and yeast [<a href="/books/n/sef/A727/?report=reader#A915">187</a>,<a href="/books/n/sef/A727/?report=reader#A1277">549</a>,<a href="/books/n/sef/A727/?report=reader#A1609">881</a>].
Certain organisms, such as <i>Euglena</i>, seem to have enzymes of
both classes. Although these two enzyme forms have similar structures, they
do not share any detectable sequence similarity [<a href="/books/n/sef/A727/?report=reader#A985">257</a>].</p><p>Sequence analysis of archaeal genomes and those of chlamydia showed that they
encode neither a typical class I enzyme, nor a typical class II enzyme.
Instead, chlamydia and all archaea, with the exception of thermoplasmas,
encode orthologs of the recently described class I aldolase DhnA (FbaB) of
<i>E. coli</i>, which is only distantly related to the regular
class I enzymes and may be considered a third class of aldolases. Recently,
fructose-1,6-bisphosphate aldolase activity was demonstrated in the
<i>P. furiosus</i> homolog of DhnA; this enzyme has been
referred to as a class IA aldolase [<a href="/books/n/sef/A727/?report=reader#A1498">770</a>]. The phyletic patterns of the bacterial-type class II
aldolase (COG0191) and the DhnA-type aldolase (COG1830) are almost
complementary, except that both types of aldolases are present in <i>E.
coli</i> and <i>A. aeolicus</i>, and none of them is
detectable in <i>X. fastidiosa</i> (<a class="figpopup" href="/books/NBK20266/figure/A378/?report=objectonly" target="object" rid-figpopup="figA378" rid-ob="figobA378">Figure 7.1</a>). <i>X. fastidiosa</i>, a plant
pathogen, encodes a eukaryotic class I aldolase, which is specifically
similar to the plant class I aldolase and probably has been acquired from
the plant host via HGT. However, typical eukaryotic (class I)
fructose-1,6-bisphosphate aldolase is also encoded in several other
bacteria, in which cases the underlying evolutionary scenario is less
clear.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA378" co-legend-rid="figlgndA378"><a href="/books/NBK20266/figure/A378/?report=objectonly" target="object" title="Figure 7.1" class="img_link icnblk_img figpopup" rid-figpopup="figA378" rid-ob="figobA378"><img class="small-thumb" src="/books/NBK20266/bin/ch7f1.gif" src-large="/books/NBK20266/bin/ch7f1.jpg" alt="Figure 7.1. Distribution of glycolysis (Embden-Meyerhoff-Parnas pathway) enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA378"><h4 id="A378"><a href="/books/NBK20266/figure/A378/?report=objectonly" target="object" rid-ob="figobA378">Figure 7.1</a></h4><p class="float-caption no_bottom_margin">Distribution of glycolysis (Embden-Meyerhoff-Parnas pathway)
enzymes in organisms with completely sequenced genomes. Each rounded rectangle shows a glycolytic enzyme, denoted by its
gene name and the COG number. Alternative enzymes catalyzing the
same <a href="/books/NBK20266/figure/A378/?report=objectonly" target="object" rid-ob="figobA378">(more...)</a></p></div></div><p>Although most genomes encode only one type of fructose-1,6-bisphosphate
aldolase, different forms of this enzyme do coexist in several organisms. In
particular, the relatively large genome of the plant symbiont <i>M.
loti</i> encodes fructose-1,6-bisphosphate aldolases of all three
classes.</p><p>The nature of the aldolase, if any, in thermoplasmas remains unclear. The
apparent absence in these archaea of both phosphofructokinase and
fructose-1,6-bisphosphate aldolase might indicate that these organisms split
hexoses into trioses exclusively via the Entner-Doudoroff pathway (see
below). Indeed, thermoplasmas encode close homologs of the recently
described fructose-6-phosphate aldolase [<a href="/books/n/sef/A727/?report=reader#A1486">758</a>].</p><p>Finally, given that chlamydiae are important human pathogens and that the
unusual class IA fructose-1,6-bisphosphate aldolase is the only aldolase
encoded in their genomes, this presumably essential enzyme might be a
promising target for anti-chlamydial drug therapy (see <a href="#A501">7.6</a>) [<a href="/books/n/sef/A727/?report=reader#A985">257</a>,<a href="/books/n/sef/A727/?report=reader#A994">266</a>].</p></div><div id="A379"><h4>Triose phosphate isomerase (EC 5.3.1.1)</h4><p>Triose phosphate isomerase is conserved in all organisms, with the exception
of <i>Rickettsia</i>. Bacterial-eukaryotic and archaeal isomerases
form two clearly separated clusters [<a href="/books/n/sef/A727/?report=reader#A967">239</a>]. This gave rise to the notion that eukaryotic triose
phosphate isomerases originated from the promitochondrial endosymbiont whose
genes have been transferred into the nucleus of the eukaryotic host [<a href="/books/n/sef/A727/?report=reader#A1160">432</a>].</p></div><div id="A380"><h4>Glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12)</h4><p>Like triosephosphate isomerases, archaeal glyceraldehyde-3-phosphate
dehydrogenases are homologous to those from bacteria and eukaryotes but form
a well-defined cluster, suggesting the mitochondrial origin of this enzyme
in eukaryotes. In pyrococci and, probably, in several other archaea, the
main glycolytic flow goes through a different enzyme,
glyceraldehyde-3-phosphate:ferredoxin oxidoreductase, whereas
glyceraldehyde-3-phosphate dehydrogenase appears to be confined to
gluconeogenesis [<a href="/books/n/sef/A727/?report=reader#A1312">584</a>,<a href="/books/n/sef/A727/?report=reader#A1595">867</a>].</p><p>In <i>U. urealyticum</i>, the typical NADH-dependent
glyceraldeldehyde-3-phosphate dehydrogenase is missing, and this reaction is
apparently catalyzed by a non-phosphorylating, NADP-dependent enzyme,
similar to the well-characterized enzymes from plants and
<i>Streptococcus mutans</i> [<a href="/books/n/sef/A727/?report=reader#A1054">326</a>,<a href="/books/n/sef/A727/?report=reader#A1272">544</a>]. These enzymes
belong to a large superfamily of NADP-dependent aldehyde dehydrogenases and
are unrelated to the phosphorylating glyceraldeldehyde-3-phosphate
dehydrogenase [<a href="/books/n/sef/A727/?report=reader#A1296">568</a>]. Remarkably, an
archaeal member of the non-phosphorylating glyceraldeldehyde-3-phosphate
dehydrogenase family uses NAD instead of NADP [<a href="/books/n/sef/A727/?report=reader#A852">124</a>].</p></div><div id="A381"><h4>Phosphoglycerate kinase (EC 2.7.2.3)</h4><p>Like triose phosphate isomerases and glyceraldehyde-3-phosphate
dehydrogenases, phosphoglycerate kinase is conserved in all organisms that
have glycolysis, and the sequences from bacteria and eukaryotes are closer
to each other than they are to their archaeal counterparts, suggesting the
mitochondrial origin of the eukaryotic enzyme.</p></div><div id="A382"><h4>Phosphoglycerate mutase (EC 5.4.2.1)</h4><p>The diversity of phosphoglycerate mutases was discussed earlier (see <a href="/books/n/sef/A22/?report=reader#A43">2.2.6</a>). We would only like to reiterate
that there are two unrelated forms of this enzyme,
2,3-bisphosphoglycerate-dependent (animal-type) and
2,3-bisphosphoglycerate-independent (plant-type), either one of which (or
both) can be found in various bacteria [<a href="/books/n/sef/A727/?report=reader#A866">138</a>]. Although <i>E. coli pgm</i> mutants devoid of
its principal (cofactor-dependent) form of phosphoglycerate mutase clearly
exhibit a mutant phenotype, a recent study of the second
(cofactor-independent) form of this enzyme showed that it accounts for as
much as 10% of the total phosphoglycerate mutase activity in
<i>E. coli</i> [<a href="/books/n/sef/A727/?report=reader#A972">244</a>].</p><p>Remarkably, neither form of phosphoglycerate mutase is encoded in any
archaeal genome available to date, with the sole exception of
<i>Halobacterium</i> spp., which has a typical
cofactor-independent enzyme, similar to the one in <i>B.
subtilis</i>. Sequence analysis of archaeal genomes showed that
they encode enzymes of the alkaline phosphatase superfamily that are
distantly related to the cofactor-independent phosphoglycerate mutase and
contain all the principal active-site residues [<a href="/books/n/sef/A727/?report=reader#A986">258</a>]. These enzymes were predicted to have a
phosphoglycerate mutase activity [<a href="/books/n/sef/A727/?report=reader#A986">258</a>]. This prediction was supported by the structural analysis
of the cofactor-independent phosphoglycerate mutase [<a href="/books/n/sef/A727/?report=reader#A989">261</a>,<a href="/books/n/sef/A727/?report=reader#A1122">394</a>] and
has been recently confirmed by direct experimental data [<a href="/books/n/sef/A727/?report=reader#A1036">308</a>,<a href="/books/n/sef/A727/?report=reader#A1594">866</a>]. Thus, like phosphofructokinase and
fructose-1,6-bisphosphate aldolase, phosphoglycerate mutase is found in
three different (unrelated or distantly related) variants.</p></div><div id="A383"><h4>Enolase (EC 4.2.1.11)</h4><p>Enolases encoded in bacterial, archaeal, and eukaryotic genomes are highly
conserved; phylogenetic trees for enolases show a &#x0201c;star
topology&#x0201d;, which precludes any definitive conclusions on the
evolutionary scenario for this enzyme. Pyrococci and <i>M.
jannaschii</i> encode additional, divergent paralogs of enolase
whose function(s) remains unknown.</p></div><div id="A384"><h4>Pyruvate kinase (EC 2.7.1.40)</h4><p>Pyruvate kinase, the terminal glycolytic enzyme, is not encoded in some
bacterial (<i>A. aeolicus</i>, <i>T. pallidum</i>) and
archaeal (<i>A. fulgidus</i>, <i>M.
thermoautotrophicum</i>) genomes. In these organisms, the pyruvate
kinase function is probably taken over by phosphoenolpyruvate synthase,
which is capable of catalyzing pyruvate formation by reversing its typical
reaction.</p><p>Pyruvate kinase, like phosphofructokinase (see above), is also missing in
<i>H. pylori</i>. Although a ribokinase-like
phosphofructokinase and phosphoenolpyruvate synthase could be considered as
possible bypasses for these enzymes, it seems more likely that glycolysis is
not functional in <i>H. pylori.</i> In contrast, this bacterium
encodes the complete set of enzymes involved in gluconeogenesis (<a href="/books/n/sef/A517/?report=reader#A523">Figure 8.2</a>). Such organization of
metabolism seems to make perfect sense for <i>H. pylori</i>, given
the challenge of maintaining near-neutral intracellular pH in the highly
acidic gastric environment. Sugar fermentation, resulting in intracellular
production of acid, would place an additional burden on the pH maintenance
mechanism, whereas gluconeogenesis converts organic acids into sugars and
thus removes H from the cytoplasm. For the purposes of energy production,
<i>H. pylori</i> apparently depends on fermentation of amino
acids and oligopeptides that are produced by gastric proteolysis and are
transported into the bacterial cells by ABC-type transporters. Amino acid
fermentation results in alkalinization of the cytoplasm and could relieve
part of the burden of pH maintenance in <i>H. pylori</i>. This
simple example shows that, even when seemingly plausible candidates for
missing steps in a pathway can be suggested, this should be done with
caution, and the resulting predicted pathways should be assessed against the
biological background of the respective organism.</p><p>After a string of recent publications [<a href="/books/n/sef/A727/?report=reader#A1059">331</a>,<a href="/books/n/sef/A727/?report=reader#A1111">383</a>,<a href="/books/n/sef/A727/?report=reader#A1498">770</a>], it appears that most glycolytic
enzymes have now been accounted for. While there are no clear candidates for
phosphofructokinase and fructose-1,6-bisphosphate aldolase in
<i>Thermoplasma</i> spp., the chances of discovering new
enzyme variants in this pathway appear very slim.</p></div></div><div id="A385"><h3>7.1.2. Gluconeogenesis</h3><p>With the exception of reactions catalyzed by phosphofructokinase and pyruvate
kinase, glycolytic reactions are reversible and function also in gluconeogenesis
(<a class="figpopup" href="/books/NBK20266/figure/A386/?report=objectonly" target="object" rid-figpopup="figA386" rid-ob="figobA386">Figure 7.2</a>). The reversal of the
latter reaction, i.e. conversion of pyruvate into phosphoenolpyruvate, can be
catalyzed by two closely related enzymes, phosphoenolpyruvate synthase and
pyruvate, phosphate dikinase. The only other reaction that is specific for
gluconeogenesis is the dephosphorylation of fructose-1,6-bisphosphate.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA386" co-legend-rid="figlgndA386"><a href="/books/NBK20266/figure/A386/?report=objectonly" target="object" title="Figure 7.2" class="img_link icnblk_img figpopup" rid-figpopup="figA386" rid-ob="figobA386"><img class="small-thumb" src="/books/NBK20266/bin/ch7f2.gif" src-large="/books/NBK20266/bin/ch7f2.jpg" alt="Figure 7.2. Distribution of gluconeogenesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA386"><h4 id="A386"><a href="/books/NBK20266/figure/A386/?report=objectonly" target="object" rid-ob="figobA386">Figure 7.2</a></h4><p class="float-caption no_bottom_margin">Distribution of gluconeogenesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div><div id="A387"><h4>Phosphoenolpyruvate synthase (EC 2.7.9.2)</h4><p>Phosphoenolpyruvate synthase (pyruvate, water dikinase, EC 2.7.9.2) and
pyruvate, phosphate dikinase (EC 2.7.9.1) catalyze two similar reactions of
phosphoenolpyruvate biosynthesis</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e1.jpg" alt="Image ch7e1.jpg" /></div><p>and have highly similar sequences. This enzyme is widely present in bacteria,
archaea, protists, and plants but is missing in animals, where PEP is
synthesized from oxaloacetates in a PEP carboxykinase-catalyzed
reaction.</p></div><div id="A388"><h4>Phosphoenolpyruvate carboxykinase (EC 4.1.1.32 and EC 4.1.1.49)</h4><p>Phosphoenolpyruvate carboxykinase exists in two unrelated forms, which
catalyze ATP-dependent (EC 4.1.1.49) or GTP-dependent (EC 4.1.1.32)
decarboxylation of oxaloacetate:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e2.jpg" alt="Image ch7e2.jpg" /></div><p>These forms show remarkably complex phyletic distributions. The GTP-dependent
form is found in animals and in a limited number of bacteria, such as
<i>Chlamydia</i> spp., <i>Mycobacterium</i> spp.,
<i>T. pallidum</i>, and the green sulfur bacterium
<i>Chlorobium limicola</i>. Among archaea, it is encoded only
in the genomes of pyrococci, thermoplasmas, and <i>Sulfolobus</i>.
In contrast, the ATP-dependent form of phosphoenolpyruvate carboxykinase is
found in plants, yeast, and many bacteria. The only complete archaeal genome
that has been found to encode the ATP-dependent form is that of <i>A.
pernix</i> (<a class="figpopup" href="/books/NBK20266/figure/A386/?report=objectonly" target="object" rid-figpopup="figA386" rid-ob="figobA386">Figure 7.2</a>).</p><p>Since the typical bacterial ATP-dependent phosphoenolpyruvate carboxykinase
appears to be unrelated to the GTP-dependent form found in humans, this key
enzyme of central metabolism might be an interesting drug target for such
pathogenic bacteria as <i>H. influenzae</i> and <i>C.
jejuni</i>(see <a href="#A501">7.6</a>).</p><p>There have also been reports of a third, pyrophosphate-dependent, form of
phosphoenolpyruvate carboxykinase [<a href="/books/n/sef/A727/?report=reader#A1547">819</a>], but they remain unconfirmed and no sequence so far has
been identified with this form. Absent this third form, phosphoenolpyruvate
carboxykinase appears to be missing in a large number of microorganisms,
leaving room for discovery of a new enzyme.</p></div><div id="A389"><h4>Fructose-1,6-bisphosphatase (EC 3.1.3.11)</h4><p>The best-studied form of fructose-1,6-bisphosphatase, found in <i>E.
coli</i>, yeast, and human (COG0158), has a limited phyletic
distribution: it is not encoded in the genomes of chlamydia, spirochetes,
Gram-positive bacteria, <i>A. aeolicus</i>, or <i>T.
maritima</i>. Among archaea, it is present only in
<i>Halobacterium</i> sp. A second form of this enzyme
(COG1494), originally described in cyanobacteria, has been reported to
function both as a fructose-1,6-bisphosphatase and as a
sedoheptulose-1,7-bisphosphatase [<a href="/books/n/sef/A727/?report=reader#A1551">823</a>,<a href="/books/n/sef/A727/?report=reader#A1552">824</a>].</p><p>This form also has a limited phyletic distribution, being found in a
relatively small number of bacteria (<a class="figpopup" href="/books/NBK20266/figure/A386/?report=objectonly" target="object" rid-figpopup="figA386" rid-ob="figobA386">Figure
7.2</a>). Although a member of this second family is encoded in
<i>B. subtilis</i>, this organism also has a distinct form of
fructose-1,6-bisphosphatase that is unrelated to the first two and is found
only in several other low-GC, Gram-positive bacteria [<a href="/books/n/sef/A727/?report=reader#A979">251</a>]. Finally, archaea encode yet another, fourth form
of this enzyme that belongs to the inositol monophosphatase family and only
recently has been shown to possess fructose-1,6-bisphosphatase activity
[<a href="/books/n/sef/A727/?report=reader#A1127">399</a>,<a href="/books/n/sef/A727/?report=reader#A1529">801</a>]. Like <i>B. subtilis</i>, several
bacterial genomes encode members of more than one protein family, which
include known or potential fructose-1,6-bisphosphatases; this makes it hard
to predict which of them actually has this function in gluconeogenesis. In
contrast, there is no clear candidate for this function in <i>A.
aeolicus</i>, <i>T. maritima</i>, <i>X.
fastidiosa</i>, <i>Chlamydia</i> spp., mycoplasmas,
spirochetes, and thermoplasmas. While the first three of these organisms and
<i>B. burgdorferi</i> encode enzymes of the inositol
monophosphatase family, they are not closely related to the archaeal
fructose-1,6-bisphosphatase (typified by the MJ0109 protein from <i>M.
jannaschii</i>) and might represent an independent case of enzyme
recruitment. Proteins that function as fructose-1,6-bisphosphatase in
<i>Chlamydia</i> spp., <i>Thermoplasma</i> spp.,
mycoplasmas, and <i>T. pallidum</i>, if any, remain to be
identified.</p></div></div><div id="A390"><h3>7.1.3. Entner-Doudoroff pathway and pentose phosphate shunt</h3><p>Alternative pathways for converting hexoses into trioses, the pentose phosphate
shunt and the Entner-Doudoroff pathway, are found in many organisms but cannot
be considered universal. Both of these pathways start from the NADP-dependent
oxidation of glucose-6-phosphate into phosphogluconolacton and proceed through
6-phosphogluconate (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure 7.3</a>). Instead
of the standard Entner-Doudoroff pathway, some archaea encode the so-called
non-phosphorylating variant of this pathway, which starts from glucose and
includes unphosphorylated intermediates.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA391" co-legend-rid="figlgndA391"><a href="/books/NBK20266/figure/A391/?report=objectonly" target="object" title="Figure 7.3" class="img_link icnblk_img figpopup" rid-figpopup="figA391" rid-ob="figobA391"><img class="small-thumb" src="/books/NBK20266/bin/ch7f3.gif" src-large="/books/NBK20266/bin/ch7f3.jpg" alt="Figure 7.3. Distribution of enzymes of the pentose phosphate and Entner-Doudoroff pathways in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA391"><h4 id="A391"><a href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-ob="figobA391">Figure 7.3</a></h4><p class="float-caption no_bottom_margin">Distribution of enzymes of the pentose phosphate and
Entner-Doudoroff pathways in organisms with completely sequenced
genomes. Details are as in Figure
2.7. </p></div></div><div id="A392"><h4>Glucose-6-phosphate 1-dehydrogenase (EC 1.1.1.49)</h4><p>Glucose 6-phosphate dehydrogenase (Zwischenferment) primarily uses NADP as
the electron acceptor, although there have been reports of NAD-dependent
forms. This enzyme is found in many bacteria and eukaryotes but is not
encoded in any of the archaeal genomes sequenced to date. In addition, it is
missing in several bacteria, such as <i>M. leprae</i>, <i>B.
halodurans</i>, <i>S. pyogenes</i>, <i>C.
jejuni</i>, and mycoplasmas (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure
7.3</a>).</p></div><div id="A393"><h4>6-Phosphogluconolactonase (EC 3.1.1.31)</h4><p>Although this enzymatic activity had been characterized many years ago, the
gene for the lactonase remained unidentified until very recently, which was
due, in part, to the inherent instability of its substrate and, in part, to
the fact that this activity resides in a protein that is closely related to
glucosamine-6-phosphate isomerase/deaminase and might even combine both
activities [<a href="/books/n/sef/A727/?report=reader#A882">154</a>,<a href="/books/n/sef/A727/?report=reader#A1056">328</a>]. In humans, the lactonase is
fused to the glucose-6-phosphate dehydrogenase, forming the C-terminal
domain of a bifunctional enzyme. Interestingly, in <i>Plasmodium
falciparum</i>, the fusion partners switch places, with the
lactonase located at the N-terminus [<a href="/books/n/sef/A727/?report=reader#A1279">551</a>]. The lactonase is found largely in the same set of species
as glucose dehydrogenase, although it appears to be missing, additionally,
in <i>A. aeolicus</i> and <i>D. radiodurans</i>.</p></div><div id="A394"><h4>7.1.3.1. Pentose phosphate shunt</h4><div id="A395"><h5>6-Phosphogluconate dehydrogenase (decarboxylating, EC
1.1.1.44)</h5><p>6-Phosphogluconate dehydrogenase, the product of the <i>gnd</i>
gene in <i>E. coli</i>, is the upstream enzyme specific for
the pentose phosphate pathway. Of those organisms that encode
phosphogluconate dehydrogenase (COG0362), several (<i>M.
loti</i>, <i>B. subtilis</i>, <i>L.
lactis</i>) also encode its close paralog (COG1023), whose
function remains unknown but which is likely to have the same activity.
Phosphogluconate dehydrogenase has an even more narrow phylogenetic
distribution than phosphogluconolactonase, being additionally absent
from <i>S. pyogenes</i>, <i>X. fastidiosa</i>,
<i>H. pylori</i>, and <i>C. jejuni</i> (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure 7.3</a>).</p></div><div id="A396"><h5>Pentose-5-phosphate-3-epimerase (EC 5.1.3.1)</h5><p>The next reaction of the pentose phosphate pathway, isomerization of
ribulose 5-phosphate into xylulose 5-phosphate, is catalyzed by
phosphoribulose epimerase. In addition to the pentose phosphate pathway,
this enzyme also participates in the interconversions of pentose
phosphates in the Calvin cycle, which accounts for its wider phyletic
distribution than seen for phosphogluconate dehydrogenase.</p></div><div id="A397"><h5>Ribose 5-phosphate isomerase (EC 5.3.1.6)</h5><p>Ribose-5-phosphate isomerase, which catalyzes interconversion of ribulose
5-phosphate and ribose 5-phosphate, is found in two apparently unrelated
forms, both of which, RpiA and RpiB, have been characterized in
<i>E. coli</i> [<a href="/books/n/sef/A727/?report=reader#A1521">793</a>]. RpiA is found in many bacteria, archaea, and
eukaryotes. In contrast, RpiB is limited to certain bacterial species
and is the sole form of ribose-5-phosphate isomerase in <i>B.
subtilis</i>, <i>M. tuberculosis</i>, <i>H.
pylori</i>, and several other bacteria. The phyletic patterns
of the two forms of the enzyme are largely complementary:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e3.jpg" alt="Image ch7e3.jpg" /></div><p>Like phosphoribulose epimerase, phosphoribose isomerase participates in
the Calvin cycle, which might explain its universal distribution.</p></div><div id="A398"><h5>Transketolase (EC 2.2.1.1)</h5><p>In eukaryotes and bacteria, transketolase is a single protein of
610&#x02013;630 amino acid residues [<a href="/books/n/sef/A727/?report=reader#A1549">821</a>]. In archaea, however, this enzyme is either missing
altogether (e.g. <i>A. fulgidus</i>, <i>M.
thermoautotrophicum</i>) or is encoded by two separate genes
that may not even be adjacent (in <i>M. jannaschii</i>). The
hyperthermophilic bacterium <i>T. maritima</i> has both types
of genes, one full-length and one split gene, the latter probably
acquired from archaea via HGT. Transketolase shows high sequence
similarity to deoxyxylulose-5-phosphate synthase and other thiamine
pyrophosphate-dependent enzymes, which might point to a broad substrate
specificity of this enzyme, particularly in thermophiles.</p></div><div id="A399"><h5>Transaldolase (EC 2.2.1.2)</h5><p>Transaldolase is a protein of 310&#x02013;330 amino acid residues,
which is present in eukaryotes and many bacteria and catalyzes the
transfer of the tri-carbon unit of sedoheptulose-7-phosphate to
glyceraldehyde-3-phosphate, producing fructose-6-phosphate and
erythrose-4-phosphate [<a href="/books/n/sef/A727/?report=reader#A1549">821</a>].
Archaea and some other bacteria encode a closely related but shorter
protein, about 210&#x02013;230 aa long, which has recently been
demonstrated to function not as transaldolase, but as
fructose-6-phosphate aldolase, which splits fructose-6-phosphate into
glyceraldehyde-3-phosphate and dihydroxyacetone [<a href="/books/n/sef/A727/?report=reader#A1486">758</a>]. While <i>E. coli</i> encodes two
paralogous transaldolases (<i>talA</i>, <i>talB</i>)
and two paralogs of the smaller related enzyme (<i>talC</i>,
<i>mipB</i>), many other micro-prokaryotes, including
<i>B. subtilis</i>, <i>M. jannaschii</i>, and
<i>Thermoplasma</i> spp., encode only the latter protein.
Although the exact substrate specificity of these enzymes is not known,
enzymes from <i>B. subtilis</i> and <i>T.
maritima</i> have been reported to have transaldolase activity
[<a href="/books/n/sef/A727/?report=reader#A1486">758</a>]. Thus, different MipB
orthologs could have different (primary) activities, which makes
complete reconstruction of the pentose phosphate pathway in organisms
having these enzymes unrealistic at this time. Clearly, however, the
phyletic patterns of the enzymes of this pathway differ significantly,
which suggests the existence of still uncharacterized enzyme forms.</p></div></div><div id="A400"><h4>7.1.3.2. The Entner-Doudoroff pathway</h4><p>Conversion of 6-phosphogluconate into two tri-carbon molecules,
3-phosphoglyceraldehyde and pyruvate, via the Entner-Doudoroff pathway
includes only two steps, which are catalyzed by 6-phosphogluconate
dehydratase and 2-keto-3-deoxy-6-phosphogluconate aldolase (the products of
<i>E. coli</i> genes <i>edd</i> and
<i>eda</i>, respectively (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure 7.3</a>).</p><div id="A401"><h5>Phosphogluconate dehydratase (EC 4.2.1.12)</h5><p>Phosphogluconate dehydratase is a close paralog of dihydroxyacid
dehydratase, an enzyme of isoleucine/valine biosynthesis, which is
encoded in almost every genome. As a result, it is not easy to decide
which organisms encode phosphogluconate dehydratase. In <i>E.
coli</i> and several other proteobacteria, <i>edd</i>
and <i>eda</i> genes form operons. In other organisms, such as
<i>P. aeruginosa</i>, even though both these genes are
present, they are not adjacent, which complicates the identification of
phosphogluconate dehydratase.</p></div><div id="A402"><h5>2-Keto-3-deoxy-6-phosphogluconate aldolase (EC 4.1.2.14)</h5><p>KDPG aldolase has a much more narrow phyletic distribution than
phosphogluconate/dihydroxyacid dehydratase (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Fig 7.3</a>). Assuming that a functional
Entner-Doudoroff pathway requires the presence of each of these enzymes,
as well as glucose-6-phosphate dehydrogenase and
phosphogluconolactonase, the available genomic data suggest that the
pathway is limited to certain proteobacteria, <i>T.
maritima</i>, and some Gram-positive bacteria of the
Bacillus/Clostridium group.</p></div></div><div id="A403"><h4>7.1.3.3. Non-phosphorylated variants of the Entner-Doudoroff pathway</h4><p>While the standard Entner-Doudoroff pathway starts from glucose-6-phosphate
and proceeds through phosphorylated sugar intermediates, a variety of
bacteria and archaea possess so-called
&#x0201c;non-phosphorylated&#x0201d; variants of this pathway, which all
start from glucose and delay phosphorylation until later stages. The
simplest version of such a modified pathway includes glucose oxidation into
gluconate, followed by its phosphorylation into 6-phosphogluconate. The
resulting 6-phosphogluconate rejoins the standard Entner-Doudoroff pathway.
Another variant of the modified pathway includes an additional
non-phosphorylated step, dehydratation of gluconate into
2-keto-3-deoxygluconate, followed by its phosphorylation. In yet another
variant of this pathway, phosphorylation is delayed even further, until
after splitting of 2-keto-3-deoxygluconate into two tri-carbon molecules,
pyruvate and glyceraldehyde. The latter compound is then phosphorylated into
3-phosphoglyceraldehyde. Finally, phosphorylation can be delayed one step
further, with glyceraldehyde first oxidized into glycerate and then
phosphorylated into 2-phosphoglycerate.</p><div id="A404"><h5>Glucose 1-dehydrogenase (EC 1.1.1.47, 1.1.99.10)</h5><p>Glucose dehydrogenase, which catalyzes glucose oxidation into
glucono-1,5-lactone, is known in several variants, which use different
electron acceptors. Two non-orthologous NAD-dependent variants of this
enzyme (EC 1.1.1.47), typified by enzymes from <i>T.
acidophilum</i> [<a href="/books/n/sef/A727/?report=reader#A1125">397</a>]
and <i>Bacillus megaterium</i> [<a href="/books/n/sef/A727/?report=reader#A1657">929</a>], belong, respectively, to the Zn-containing
dehydrogenase family and to the short-chain reductases/dehydrogenases
family. One more variant of glucose dehydrogenase, which is present in
<i>E. coli</i> and several other bacteria, uses
pyrroloquinoline quinone as the electron acceptor [<a href="/books/n/sef/A727/?report=reader#A1365">637</a>]. Finally, the enzyme from
<i>Drosophila</i> (DHGL_DROME) is a flavoprotein that can
use a variety of electron acceptors [<a href="/books/n/sef/A727/?report=reader#A869">141</a>].</p></div><div id="A405"><h5>Gluconolactonase (EC 3.1.1.17)</h5><p>Only a single variant of gluconolactonase has been characterized so far
[<a href="/books/n/sef/A727/?report=reader#A1141">413</a>]. It has a patchy and
relatively narrow phyletic distribution (COG3386), suggesting that
alternative versions of this enzyme might exist.</p></div><div id="A406"><h5>Gluconate kinase (EC 2.7.1.12)</h5><p>Gluconate kinase is found in two distinct versions, one unique and the
other belonging to a large family of sugar kinases. This second form of
gluconate kinase has probably evolved from a glycerol kinase or a
xylulose kinase via enzyme recruitment (<a href="/books/n/sef/A22/?report=reader#A40">2.2.5</a>). Gluconate kinases of the first type are found in
yeast, <i>D. radiodurans</i>, <i>E. coli</i>, and
several other proteobacteria, whereas the second form is apparently
limited to <i>B. subtilis</i> and a handful of other
Gram-positive bacteria.</p></div><div id="A407"><h5>Gluconate dehydratase (EC 4.2.1.39)</h5><p>Although gluconate dehydratase activity has been described in bacteria
long ago [<a href="/books/n/sef/A727/?report=reader#A1027">299</a>] and can be
easily detected in archaea [<a href="/books/n/sef/A727/?report=reader#A1126">398</a>], the gene(s) for this enzyme has not been identified.
<i>E. coli</i>, some other bacteria, and
<i>Thermoplasma</i> spp. encode an enzyme with similar
activity, D-mannonate dehydratase (EC 4.2.1.8, the product of
<i>uxuA</i> gene), which converts mannonate into
2-keto-3-deoxygluconate. It is not known whether this enzyme can use
gluconate as a substrate. In any case, its narrow phyletic distribution
suggests that, even if UxuA functions as gluconate dehydratase in
<i>E. coli</i>, <i>M. loti</i>, <i>B.
subtilis</i>, and <i>Thermoplasma</i> spp., there
should exist a different form of this enzyme, which would participate in
the non-phosphorylated Entner-Doudoroff pathway in other archaea.</p></div><div id="A408"><h5>2-Keto-3-deoxygluconate aldolase</h5><p>Although splitting of 2-keto-3-deoxygluconate into pyruvate and
glyceraldehyde has been described long ago [<a href="/books/n/sef/A727/?report=reader#A744">16</a>], the first gene for 2-keto-3-deoxygluconate
has been identified only recently in the hyperthermophilic crenarchaeon
<i>S. solfataricus</i>. This enzyme is closely related to
N-acetyl-neuraminate lyase and belongs to the same superfamily of Schiff
base-dependent aldolases [<a href="/books/n/sef/A727/?report=reader#A854">126</a>].
Enzymes of this family (COG0329) are present in all archaeal genomes
sequenced so far, as well as in most bacteria. Although the exact
substrate specificity of each particular member of this family is not
yet clear, <i>Thermoplasma</i> spp. and <i>P.
abyssi</i> encode proteins that are highly similar to the
enzyme from <i>Sulfolobus</i> and can be confidently predicted
to catalyze this reaction.</p></div></div></div><div id="A409"><h3>7.1.4. The TCA cycle</h3><p>The tricarboxylic acid cycle (Krebs cycle) is the central metabolic pathway that
links together carbohydrate, amino acid, and fatty acid degradation and supplies
precursors for various biosynthetic pathways. Remarkably, the complete TCA
cycle, which has been studied in much detail in animal and yeast mitochondria,
<i>E. coli</i>, and <i>B. subtilis</i>, is only found in
a handful of microorganisms (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure 7.4</a>).
Most organisms with completely sequenced genomes encode only a certain subset of
TCA cycle enzymes and, instead of performing the entire cycle, utilize only
fragments of it. Another remarkable feature is the diversity of this pathway:
cases of non-orthologous gene displacement are detectable for at least five of
the eight TCA cycle enzymes. A detailed analysis of the phyletic distribution
and evolution of the TCA cycle enzymes has been recently published by Huynen and
coworkers [<a href="/books/n/sef/A727/?report=reader#A1098">370</a>]. Most of their
conclusions remain valid, although the sequences of the genomes of two aerobic
archaea, the crenarchaeon <i>A. pernix</i> and the euryarchaeaon
<i>Halobacterium</i> sp., have substantially changed the notions
of what can and cannot be found in archaeal genomes. In an impressive
confirmation of early biochemical results on halobacterial metabolism [<a href="/books/n/sef/A727/?report=reader#A736">8</a>], both of these organisms have been
found to encode the complete set of TCA cycle enzymes as was the microaerophile
<i>Thermoplasma</i> spp. A reconstruction of the TCA cycle
reactions occurring in each organism can be a very interesting project, which we
recommend the readers to do on their own (see <a href="/books/n/sef/A646/?report=reader">Problems</a>). We concentrate here exclusively on the cases of
non-orthologous gene displacement.</p><div id="A410"><h4>Citrate synthase (EC 4.1.3.7)</h4><p>Citrate synthase is a highly conserved enzyme, which is encoded in most
bacterial, archaeal, and eukaryotic genomes (<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure 7.3</a>). It serves as the principal port of entry of
acetyl-CoA into the TCA cycle and, in eukaryotes, is exclusively located in
the mitochondria. A very similar reaction is catalyzed by ATP:citrate lyase
(EC 4.1.3.8), which contains a citrate synthase-like domain at its
C-terminus.</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e4.jpg" alt="Image ch7e4.jpg" /></div><p>However, ATP:citrate lyase so far has been found exclusively in eukaryotes,
where it localizes in the cytoplasm and preferentially catalyzes the reverse
reaction, citrate cleavage.</p><p>Citrate synthase is missing in spirochetes and mycoplasmas, which do not
encode any enzymes of the TCA cycle. It is also missing in pyrococci,
<i>M. jannaschii</i>, <i>S. pyogenes</i>, and
<i>H. influenzae</i>, which encode unlinked branches of the
TCA cycle (<a class="figpopup" href="/books/NBK20266/figure/A411/?report=objectonly" target="object" rid-figpopup="figA411" rid-ob="figobA411">Figure 7.4</a>). It has been
suggested that the TCA cycle has evolved from two separate reductive
branches [<a href="/books/n/sef/A727/?report=reader#A1439">711</a>], which were
subsequently linked by (i) citrate synthase and (ii) either an
&#x003b1;-ketoglutarate dehydrogenase or an
&#x003b1;-ketoglutarate:ferredoxin oxidoreductase [<a href="/books/n/sef/A727/?report=reader#A1173">445</a>,<a href="/books/n/sef/A727/?report=reader#A1266">538</a>]. In
any case, due to the absence of known displacements, citrate synthase seems
to be a good indicator of the presence of a (nearly) complete TCA cycle in a
given organism.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA411" co-legend-rid="figlgndA411"><a href="/books/NBK20266/figure/A411/?report=objectonly" target="object" title="Figure 7.4" class="img_link icnblk_img figpopup" rid-figpopup="figA411" rid-ob="figobA411"><img class="small-thumb" src="/books/NBK20266/bin/ch7f4.gif" src-large="/books/NBK20266/bin/ch7f4.jpg" alt="Figure 7.4. Distribution of the TCA cycle enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA411"><h4 id="A411"><a href="/books/NBK20266/figure/A411/?report=objectonly" target="object" rid-ob="figobA411">Figure 7.4</a></h4><p class="float-caption no_bottom_margin">Distribution of the TCA cycle enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A412"><h4>Aconitase (EC 4.2.1.3)</h4><p>There are two distantly related, paralogous aconitases, referred to as
aconitase A and aconitase B, both of which are present in <i>E.
coli</i> and many other proteobacteria (<a class="figpopup" href="/books/NBK20266/figure/A411/?report=objectonly" target="object" rid-figpopup="figA411" rid-ob="figobA411">Figure 7.4</a>). Aconitase A has a much wider phyletic
distribution and is the form of the enzyme present in
&#x003b1;-proteobacteria <i>M. loti</i>, <i>C.
crescentus</i>, and <i>R. prowazekii</i>. Accordingly,
this is also the form of aconitase found in the mitochondria. Although
aconitase B has a much more narrow phyletic distribution, it is the only
form of the enzyme encoded in <i>Synechocystis</i> sp., <i>P.
multocida</i>, <i>H. pylori</i>, and <i>C.
jejuni</i>. </p><p>Aconitase is closely related to 3-isopropylmalate dehydratase, an enzyme of
leucine biosynthesis (see <a href="#A484">7.4.4</a>),
which sometimes makes its annotation of these enzymes in sequenced genomes
not entirely straightforward. However, <i>leu</i> genes are
usually found in a conserved operon, which helps make the correct
assignment.</p></div><div id="A413"><h4>Isocitrate dehydrogenase (EC 1.1.1.42)</h4><p>Like aconitase, isocitrate dehydrogenase is also found in two forms, which,
however, appear to be unrelated. The mitochondrial form of this enzyme, also
found in <i>E. coli</i> and many other bacteria and archaea, is
closely related to isopropylmalate dehydrogenase, an enzyme of leucine
biosynthesis (see <a href="#A484">7.4.4</a>), and it is
believed that it could have evolved via a duplication of the
<i>leuB</i> gene [<a href="/books/n/sef/A727/?report=reader#A1307">579</a>]. Again, genome annotation here has to rely on the genetic
context, i.e. on the presence or absence of adjacent <i>leu</i>
genes. In any case, the product of such a gene is very likely to have both
isocitrate dehydrogenase and isopropylmalate dehydrogenase activity. This
form is active as a homodimer, which distinguishes it from the second form,
referred to as monomeric isocitrate dehydrogenase. This second form was
originally found in <i>Vibrio</i> sp. [<a href="/books/n/sef/A727/?report=reader#A1542">814</a>] and was subsequently discovered in many other
bacteria [<a href="/books/n/sef/A727/?report=reader#A1531">803</a>]. It is the only form
of the enzyme encoded in the genomes of <i>M. leprae</i>,
<i>V. cholerae</i>, and <i>C. jejuni</i>.</p></div><div id="A414"><h4>2-Ketoglutarate dehydrogenase (EC 1.2.4.2)</h4><p>In mitochondria and many aerobic bacteria and archaea, decarboxylation of
&#x003b1;-ketoglutarate into the succinyl moiety of succinyl-CoA is
catalyzed by the thiamine pyrophosphate and lipoate-dependent
&#x003b1;-ketoglutarate dehydrogenase complex. In contrast, many anaerobic
bacteria and archaea utilize &#x003b1;-ketoglutarate ferredoxin
oxidoreductase, an unrelated enzyme [<a href="/books/n/sef/A727/?report=reader#A1173">445</a>,<a href="/books/n/sef/A727/?report=reader#A1266">538</a>].</p></div><div id="A415"><h4>Succinyl-CoA synthetase (EC 6.2.1.4, 6.2.1.5)</h4><p>Succinyl-CoA synthetases are divided into paralogous, highly similar
GTP-dependent and ATP dependent forms. Succinyl-CoA synthetase is a member
of a large family of acyl-CoA synthetases (NDP-forming), which also includes
acetyl-CoA synthetase found in many archaea and lower eukaryotes. For ATP
binding, these enzymes employ the ATP-grasp domain (<a href="/books/n/sef/A55/?report=reader#A100">Table 3.2</a>). Variants of this enzyme with shifted
substrate specificities are found in most phylogenetic lineages.</p></div><div id="A416"><h4>Succinate dehydrogenase/fumarate reductase (EC 1.3.99.1)</h4><p>Mitochondrial succinate dehydrogenase, which couples the oxidation of
succinate to fumarate with the reduction of ubiquinone to ubiquinol,
consists of four subunits carrying three iron-sulfur centers, a covalently
bound flavin and two b-type hemes (the history of the discovery of these
complexes is vividly described in [<a href="/books/n/sef/A727/?report=reader#A1501">773</a>]). The fumarate reductase (quinol:fumarate reductase)
complex also contains iron-sulfur centers and a covalently bound flavin but
usually consists of only two or three subunits. Succinate dehydrogenase is
part of the aerobic respiratory chain, whereas fumarate reductase is
involved in anaerobic respiration, with fumarate functioning as the terminal
electron acceptor. Accordingly, one or both of these enzymes is found in all
organisms, with the exception of pyrococci, spirochetes, and mycoplasmas
(<a class="figpopup" href="/books/NBK20266/figure/A411/?report=objectonly" target="object" rid-figpopup="figA411" rid-ob="figobA411">Figure 7.4</a>).</p></div><div id="A417"><h4>Fumarate hydratase (fumarase, EC 4.2.1.2)</h4><p>Like several other TCA cycle enzymes, fumarase is represented by two
unrelated forms. The mitochondrial form of this enzyme (class II) is also
encoded in many bacteria and in aerobic archaea, <i>A. pernix</i>
and <i>Halobacterium</i> sp. The second form of fumarase (class I)
consists of two subunits that are fused in most bacterial genomes but are
encoded by separate genes in archaea, <i>T. maritima</i> and
<i>A. aeolicus</i>. The two forms of fumarase have largely
complementary phyletic patterns:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e5.jpg" alt="Image ch7e5.jpg" /></div><p>The only archaeal genome that appears not to encode a fumarase is <i>T.
acidophilum</i>, whose fumarase homolog Ta0258 is much more closely
related to aspartate ammonia-lyase (COG1027) than to a typical fumarase
(COG0114). The actual activity of this <i>Thermoplasma</i> enzyme
has not been determined.</p></div><div id="A418"><h4>Malate dehydrogenase (EC 1.1.1.37)</h4><p>Malate dehydrogenase is also found in two forms, with the mitochondrial form
showing a much wider phyletic distribution. The second form of malate
dehydrogenase was originally described in archaea [<a href="/books/n/sef/A727/?report=reader#A736">8</a>,<a href="/books/n/sef/A727/?report=reader#A806">78</a>,<a href="/books/n/sef/A727/?report=reader#A1048">320</a>] and is often referred to as the
&#x0201c;archaeal&#x0201d; form of the enzyme. However, it is also
encoded in certain bacterial genomes, including three paralogous genes in
<i>E. coli</i> (<i>ybiC</i>, <i>yiaK</i>,
and <i>ylbC</i>) and <i>M. loti</i>. It is the only form
of malate dehydrogenase in pyrococci and in <i>P. aeruginosa</i>.
Remarkably, <i>M. thermoautotrophicus, M. jannaschii, B. subtilis, H.
influenzae</i>, and <i>P. multocida</i> encode both forms
of malate dehydrogenase ([<a href="/books/n/sef/A727/?report=reader#A1570">842</a>],
<a class="figpopup" href="/books/NBK20266/figure/A391/?report=objectonly" target="object" rid-figpopup="figA391" rid-ob="figobA391">Figure 7.3</a>). Why do these
organisms, with their relatively small genomes, need two paralogous forms of
this enzyme remains unclear. <i>U. urealyticum</i> and <i>T.
pallidum</i> do not encode either of the two forms of malate
dehydrogenase, in contrast to their respective relatives <i>M.
genitalium</i> and <i>B. burgdorferi</i>. Therefore, the
possibility remains that there exists yet another, third form of this
enzyme.</p></div></div></div><div id="A419"><h2 id="_A419_">7.2. Pyrimidine Biosynthesis</h2><p>In contrast to the pathways of carbohydrate metabolism discussed above, enzymes of
the pyrimidine biosynthesis pathway show a fairly consistent phyletic pattern,
although cases of non-orthologous gene displacement can be found here, too (see
<a href="/books/n/sef/A22/?report=reader#A35">Figure 2.7</a>
). The whole pathway, with the exception of the last three steps,
is missing in the obligate parasitic bacteria with small genomes: rickettsiae,
chlamydiae, spirochetes, and mycoplasmas, whereas bacteria and archaea with larger
genomes encode all or almost all enzymes of pyrimidine biosynthesis.</p><div id="A420"><h3>Carbamoyl phosphate synthase (EC 6.3.5.5)</h3><p>In bacteria and archaea, carbamoyl phosphate synthase consists of two subunits,
which in eukaryotes are fused into a single multifunctional CAD protein that
additionally contains dihydroorotase and aspartate carbamoyl-transferase
domains. The small subunit, encoded by the <i>carA</i> gene, is a
typical glutamine amidotransferase of the Triad family [<a href="/books/n/sef/A727/?report=reader#A1664">936</a>]. The large subunit consists of two ATP-grasp domains
(see <a href="/books/n/sef/A55/?report=reader#A98">3.3.3</a>) fused in the same polypeptide
chain [<a href="/books/n/sef/A727/?report=reader#A1119">391</a>,<a href="/books/n/sef/A727/?report=reader#A1528">800</a>,<a href="/books/n/sef/A727/?report=reader#A1569">841</a>]. In
<i>M. jannaschii</i> and <i>M. thermoautotrophicus</i>,
the large subunit is split into two proteins, which are encoded by different,
albeit adjacent, genes. In addition to the obligate parasites mentioned above,
carbamoyl phosphate synthase is missing in <i>P. horikoshii</i>,
<i>P. abyssi</i>, <i>Thermoplasma</i> spp., and
<i>H. influenzae</i> (<a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>). It is present, however, in <i>Pyrococcus
furiosus</i>, suggesting a relatively recent loss of this enzyme in the
other two pyrococci. In <i>P. abyssi</i>, carbamoyl phosphate
biosynthesis is carried out by an unrelated form of the enzyme, which is closely
related to carbamate kinase [<a href="/books/n/sef/A727/?report=reader#A1411">683</a>,<a href="/books/n/sef/A727/?report=reader#A1412">684</a>]. This second form is also
responsible for the carbamoyl phosphate synthase activity in <i>P.
furiosus</i> [<a href="/books/n/sef/A727/?report=reader#A929">201</a>,<a href="/books/n/sef/A727/?report=reader#A1420">692</a>] and might account for this activity
in <i>Thermoplasma</i> spp. Although both subunits of carbamoyl
phosphate synthase belong to large protein superfamilies and are similar to many
proteins with different substrate specificities, the sheer size of the large
subunit, which typically contains more than 1,050 amino acid residues, allows an
easy identification of this enzyme in genome analyses. However, caution is due
with respect to the annotation of any shorter proteins that give statistically
significant hits to the large subunit of carbamoyl phosphate synthase: these are
likely to be other ATP-grasp superfamily enzymes (see <a href="/books/n/sef/A55/?report=reader#A100">Table 3.2</a>).</p></div><div id="A421"><h3>Aspartate carbamoyltransferase (EC 2.1.3.2)</h3><p>Aspartate carbamoyltransferase, the second enzyme of pyrimidine biosynthesis, has
a wide distribution with a phyletic pattern, which is similar to that of
carbamoyl phosphate synthase but additionally includes pyrococci and
<i>Thermoplasma</i> spp. (<a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>). This enzyme, however, is lacking in <i>H.
influenzae</i> and in its close relative <i>P. multocida</i>.
In eukaryotes, aspartate carbamoyltransferase comprises the C-terminal domain of
the multifunctional CAD protein [<a href="/books/n/sef/A727/?report=reader#A1499">771</a>].</p></div><div id="A422"><h3>Dihydroorotase (EC 3.5.2.3)</h3><p>The well-characterized form of dihydroorotase (COG0418), encoded by the
<i>E. coli pyrC</i> gene [<a href="/books/n/sef/A727/?report=reader#A793">65</a>] and by the <i>URA4</i> gene in yeast [<a href="/books/n/sef/A727/?report=reader#A1053">325</a>], has a very limited phyletic
distribution (<a href="/books/n/sef/A22/?report=reader#A35">Figure 2.7</a>). In contrast,
the second form of this enzyme (COG0044) is almost universal, being present in
many bacteria, archaea, and eukaryotes [<a href="/books/n/sef/A727/?report=reader#A1414">686</a>]. In eukaryotes, this enzyme forms the middle portion of the
multifunctional CAD protein [<a href="/books/n/sef/A727/?report=reader#A1499">771</a>,<a href="/books/n/sef/A727/?report=reader#A1671">943</a>]. In yeast, however, this domain is
apparently inactive [<a href="/books/n/sef/A727/?report=reader#A1522">794</a>], most likely
because of the presence of the alternative form of dihydroorotase. Simlilar to
aspartate carbamoyltransferase, neither form of dihydroorotase is encoded in
<i>H. influenzae</i> or <i>P. multocida</i>. Notably,
the union of the phyletic patterns for the two forms of dihydroorotase is
identical to the phyletic pattern of aspartate carbamoyl-transferase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e6.jpg" alt="Image ch7e6.jpg" /></div></div><div id="A423"><h3>Dihydroorotate dehydrogenase (EC 1.3.3.1)</h3><p>Dihydroorotate dehydrogenase displays the same phyletic pattern as dihydroorotase
and aspartate carbamoyltransferase, with the addition of <i>H.
influenzae</i> and <i>P. multocida</i>. Both these bacteria
encode the enzymes for all downstream steps of pyrimidine biosynthesis.</p></div><div id="A424"><h3>Orotate phosphoribosyltransferase (EC 2.4.2.10)</h3><p>The phyletic pattern of orotate phosphoribosyltransferase differs from that of
dihydroorotate dehydrogenase in only one respect, the presence of a
<i>pyrE</i>-related gene in <i>C. pneumoniae</i>. The
function of the product of this gene in <i>C. pneumoniae</i> is
unknown, but given the absence in this organism of the enzymes for the upstream
and the downstream steps of the pathway, it is unlikely to function as orotate
phosphoribosyltransferase. Rather, this enzyme might be recruited to catalyze a
different phosphoribosyltransferase reaction. In eukaryotes, orotate
phosphoribosyltransferase is fused to the next enzyme of the pathway, OMP
decarboxylase, forming a two-domain UMP synthase. As a result, orotate
phosphoribosyltransferase and OMP decarboxylase are occasionally misannotated as
UMP synthases and vice versa [<a href="/books/n/sef/A727/?report=reader#A992">264</a>].</p></div><div id="A425"><h3>Orotidine-5&#x02032;-monophosphate decarboxylase (EC 4.1.1.23)</h3><p>Although the phyletic pattern of OMP decarboxylase is identical to that of
dihydroorotate dehydrogenase, a closer look at COG0284 shows that it consists of
three distantly related families. Two of these include well-characterized
enzymes from <i>E. coli</i> and other bacteria [<a href="/books/n/sef/A727/?report=reader#A1584">856</a>] and from yeast and other eukaryotes [<a href="/books/n/sef/A727/?report=reader#A962">234</a>,<a href="/books/n/sef/A727/?report=reader#A1298">570</a>]. The third family includes OMP decarboxylases from archaea and
a small number of bacteria, such as <i>M. tuberculosis</i>, <i>M.
leprae</i>, and <i>Myxococcus xanthus</i> [<a href="/books/n/sef/A727/?report=reader#A740">12</a>,<a href="/books/n/sef/A727/?report=reader#A1167">439</a>]. Mycobacterial OMP decarboxylases seem to be sufficiently
distinct from those of eukaryotes and other bacteria to consider them promising
targets for antituberculine drugs [<a href="/books/n/sef/A727/?report=reader#A994">266</a>].</p></div><div id="A426"><h3>Uridylate kinase (EC 2.7.4.-, 2.7.4.14)</h3><p>There seem to be two distinct forms of uridylate kinase: one specific for UMP and
found in bacteria and archaea (COG0528) and another one that phosphorylates both
UMP and CMP and is found in eukaryotes [<a href="/books/n/sef/A727/?report=reader#A1356">628</a>,<a href="/books/n/sef/A727/?report=reader#A1467">739</a>]. The eukaryotic
form of the enzyme is closely related to bacterial adenylate kinase and could
have been recruited from an ancestral prokaryotic adenylate kinase. The
prokaryotic form of uridylate kinase is encoded in all bacterial and archaeal
genomes sequenced to date, including the &#x02018;minimal&#x02019; (see
<a href="/books/n/sef/A22/?report=reader#A40">2.2.5</a>) genomes of mycoplasmas and
<i>Buchnera</i>.</p></div><div id="A427"><h3>Nucleoside diphosphate kinase (EC 2.7.4.6)</h3><p>Nucleotide diphosphate kinase (COG0105) is highly conserved in most bacteria,
archaea, and eukaryotes. Surprisingly, however, this enzyme is not encoded in
<i>T. maritima</i>, <i>L. lactis</i>, <i>S.
pyogenes</i>, and mycoplasmas. One could imagine that these organisms
employ a different nucleotide diphosphate kinase that might have been recruited,
just like the eukaryotic uridylate kinase, from the adenylate kinase family
(COG0563).</p><p>This, however, would not solve the problem for <i>T. maritima</i> and
mycoplasmas, which encode only a single enzyme of that family. It therefore
seems likely that nucleotide diphosphate kinase in these organisms has been
recruited from yet another kinase family. Indeed, a phyletic pattern search for
a protein that would be encoded in those four genomes, but not in other
organisms with relatively small genomes, such as chlamydiae, spirochetes, or
<i>H. pylori</i>, easily finds an uncharacterized (predicted)
kinase related to dihydroxyacetone kinase (COG1461), which appears to be a good
candidate for the role of nucleoside diphosphate kinase in these organisms:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e7.jpg" alt="Image ch7e7.jpg" /></div></div><div id="A428"><h3>CTP synthase (UTP-ammonia ligase, EC 6.3.4.2)</h3><p>CTP synthase is a two-domain protein, which consists of an N-terminal
nucleotide-binding synthetase domain and a C-terminal glutamine amidotransferase
domain. This enzyme is extremely highly conserved in bacteria, archaea, and
eukaryotes. It is missing only in the genomes of <i>M. genitalium</i>
and <i>M. pneumoniae</i>, which apparently make CTP from CDP or CMP in
a salvage pathway, rather than from UTP.</p><div id="A429"><h4>General notes on pyrimidine biosynthesis evolution</h4><p>Comparison of the phyletic patterns for the enzymes of pyrimidine
biosynthesis reveals two important evolutionary trends. First, there appears
to be a tendency toward decreasing the genome size by losing genes that have
ceased to be essential. Indeed, ample evidence indicates that mycoplasmas
evolved from a Gram-positive ancestor by way of massive gene loss associated
with their adaptation to parasitism. While bacilli, lactococci, and many
other Gram-positive bacteria carry the full set of genes of pyrimidine
biosynthesis, most of the <i>pyr</i> genes have been lost in the
mycoplasmal lineage. Similarly, many <i>pyr</i> genes apparently
have been lost in other parasitic bacteria with small genomes, such as
spirochetes, rickettsiae, and chlamydiae (<a href="/books/n/sef/A22/?report=reader#A35">Figure 2.7</a>).</p><p>The trend toward gene loss is much more pronounced for the initial steps of
the pyrimidine biosynthesis pathway than it is for the distal steps. Thus,
genes for the first three steps of pyrimidine biosynthesis from bicarbonate
and ammonia to dihydroorotate (<i>carA</i>, <i>carB</i>,
<i>pyrB</i>, and <i>pyrC</i>) are missing in
<i>H. influenzae</i>, but the genes for all the subsequent
steps of pyrimidine biosynthesis, from dihydroorotate to CTP, are present
(<a href="/books/n/sef/A22/?report=reader#A35">Figure 2.7</a>). This means that,
although <i>H. influenzae</i> is incapable of <i>de
novo</i> pyrimidine biosynthesis, it still can synthesize UTP and
CTP from dihydroorotate, orotate, or OMP. Spirochetes, chlamydiae,
rickettsiae, and mycoplasmas show an even deeper loss of pyrimidine
biosynthesis genes but nevertheless retain genes for the last three steps of
the pathway, the conversion of UMP into CTP. Thus, while depending on the
host for the supply of essential nutrients, this strategy allows the
parasite to preserve at least some metabolic plasticity. In particular,
every organism seems to encode enzymes to synthesize its own nucleoside
triphosphates (NTPs). For thermodynamic reasons, bacteria cannot import NTPs
directly, although intracellular bacterial parasites do encode ATP/ADP
translocases, which are capable of exchanging ADP generated by the parasite
for cytoplasmic ATP [<a href="/books/n/sef/A727/?report=reader#A1627">899</a>,<a href="/books/n/sef/A727/?report=reader#A1638">910</a>].</p></div></div></div><div id="A430"><h2 id="_A430_">7.3. Purine Biosynthesis</h2><p>Like pyrimidine biosynthesis enzymes, enzymes of the purine biosynthesis pathway
follow a consistent phylogenetic pattern, albeit with some inevitable complications
(<a class="figpopup" href="/books/NBK20266/figure/A431/?report=objectonly" target="object" rid-figpopup="figA431" rid-ob="figobA431">Figure 7.5</a>). With only a few exceptions,
enzymes that catalyze the common reactions of the pathway, which leads to the
formation of inosine-5&#x02032;-monophosphate, are missing in parasitic bacteria
with small genomes, namely mycoplasmas, rickettsiae, chlamydiae, spirochetes,
<i>Buchnera</i> sp., and <i>H. pylori</i>, and,
interestingly, in the aerobic crenarchaeon <i>A. pernix</i>. Other
bacteria encode the complete set of purine biosynthesis enzymes, whereas the
distribution of these enzymes in archaeal genomes is more complex and has to be
discussed separately for each enzyme.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA431" co-legend-rid="figlgndA431"><a href="/books/NBK20266/figure/A431/?report=objectonly" target="object" title="Figure 7.5" class="img_link icnblk_img figpopup" rid-figpopup="figA431" rid-ob="figobA431"><img class="small-thumb" src="/books/NBK20266/bin/ch7f5.gif" src-large="/books/NBK20266/bin/ch7f5.jpg" alt="Figure 7.5. Distribution of purine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA431"><h4 id="A431"><a href="/books/NBK20266/figure/A431/?report=objectonly" target="object" rid-ob="figobA431">Figure 7.5</a></h4><p class="float-caption no_bottom_margin">Distribution of purine biosynthesis enzymes in organisms with
completely sequenced genomes. All details as in Figure 2.7. </p></div></div><div id="A432"><h3>Phosphoribosylpyrophosphate synthetase (EC 2.7.6.1)</h3><p>PRPP synthetase (ribose-phosphate diphosphokinase) is an enzyme that is shared by
purine biosynthesis and histidine biosynthesis pathways. This enzyme is found in
most completely sequenced genomes, including those of mycoplasmas, spirochetes,
and <i>Buchnera</i>, which do not encode most purine biosynthesis
enzymes (<a class="figpopup" href="/books/NBK20266/figure/A431/?report=objectonly" target="object" rid-figpopup="figA431" rid-ob="figobA431">Figure 7.5</a>).</p></div><div id="A433"><h3>Amidophosphoribosyltransferase (EC 2.4.2.14)</h3><p>Glutamine phosphoribosylpyrophosphate amidotransferase (PurF) belongs to the
N-terminal nucleophile (Ntn) family of glutamine amidotransferases [<a href="/books/n/sef/A727/?report=reader#A1664">936</a>]. This enzyme is encoded in every
sequenced bacterial genome, with the exception of some obligate parasites, such
as rickettsiae, chlamydiae, spirochetes, mycoplasmas, and <i>H.
pylori,</i> and in every archaeal genome except for <i>A.
pernix</i>. The same phyletic pattern is seen for the majority of
purine biosynthesis enzymes.</p></div><div id="A434"><h3>Phosphoribosylamine-glycine ligase (EC 6.3.4.13)</h3><p>Phosphoribosylglycinamide synthetase PurD, an ATP-grasp superfamily (<a href="/books/n/sef/A55/?report=reader#A100">Table 3.2</a>) enzyme, has the same phyletic
pattern as amidophosphoribosyltransferase and many other enzymes of this
pathway.</p></div><div id="A435"><h3>Phosphoribosylglycinamide formyltransferase (EC 2.1.2.2)</h3><p>5&#x02032;-Phosphoribosyl-N-formylglycinamide synthase (GAR transformylase)
exists in two different forms, formate-dependent (PurN) and folate-dependent
(PurT), which are unrelated to each other and catalyze entirely different
reactions. The folate-dependent form functions as a transferase, catalyzing
transfer of the formyl group from formyltetrahydrofolate to
phosphoribosylglycinamide. This enzyme is found in many bacteria and eukaryotes
but only in a few archaea, such as <i>Halobacterium</i> sp. and
<i>Thermoplasma</i> spp. The formate-dependent form of the enzyme
belongs to the ATP-grasp superfamily (see <a href="/books/n/sef/A55/?report=reader#A100">Table 3.2</a>) and catalyzes an ATP-dependent ligation of
phosphoribosylglycinamide with formic acid. This is the only form of GAR
transformylase in methanogens and pyrococci. Surprisingly, neither form of the
enzyme is encoded in the <i>A. fulgidus</i> genome. With the exception
of <i>A. fulgidus</i>, the combined phyletic pattern of the two forms
of GAR transformylase coincides with the patterns for
amidophosphoribosyltransferase and phosphoribosylamine-glycine ligase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e8.jpg" alt="Image ch7e8.jpg" /></div></div><div id="A436"><h3>Phosphoribosylformylglycinamidine synthase (EC 6.3.5.3)</h3><p>Like many other amidotransferases, phosphoribosylformylglycinamidine (FGAM)
synthase PurL consists of two subunits, a glutamine amidotransferase of the
Triad family [<a href="/books/n/sef/A727/?report=reader#A1664">936</a>] and a synthetase.
The phyletic pattern of both FGAM synthase subunits is the same as that of PurF
and PurD. In <i>E. coli</i> and many other &#x003b3;-proteobacteria,
as well as in yeast and other eukaryotes, these two subunits are fused in one
polypeptide chain, whereas in most other bacteria and in archaea they are
encoded by separate genes. In this latter case, FGAM synthase apparently
requires an additional 80-aa subunit, referred to as PurS [<a href="/books/n/sef/A727/?report=reader#A1475">747</a>].</p></div><div id="A437"><h3>Phosphoribosylaminoimidazol synthetase (EC 6.3.3.1)</h3><p>Phosphoribosylformylglycinamidine cycloligase (AIR synthetase) PurM has the same
phyletic pattern as PurF, PurD, and PurL.</p></div><div id="A438"><h3>Phosphoribosylaminoimidazole carboxylase (EC 4.1.1.21)</h3><p>Phosphoribosylaminoimidazole (AIR) carboxylase (NCAIR synthetase) PurK is, like
PurD, an ATP-grasp superfamily enzyme (<a href="/books/n/sef/A55/?report=reader#A100">Table
3.2</a>), which catalyzes ATP-dependent carboxylation of AIR. Unlike
other enzymes of purine biosynthesis, PurK is not encoded in the genomes of
<i>A. fulgidus</i>, <i>C. jejuni</i>, methanogens, and
pyrococci (<a class="figpopup" href="/books/NBK20266/figure/A431/?report=objectonly" target="object" rid-figpopup="figA431" rid-ob="figobA431">Figure 7.5</a>), so that the
mechanism of AIR carboxylation in these organisms remains unknown. This reaction
can occur spontaneously at elevated temperatures in a CO<sub>2</sub>-rich
atmosphere, which could explain the absence of this enzyme in hyperthermophilic
archaea. This explanation does not seem to work, however, for <i>C.
jejuni</i>, suggesting the existence of a still unidentified
alternative version of PurK (see [<a href="/books/n/sef/A727/?report=reader#A998">270</a>,<a href="/books/n/sef/A727/?report=reader#A1295">567</a>] for
discussion).</p></div><div id="A439"><h3>Phosphoribosylcarboxyaminoimidazole mutase</h3><p>Phosphoribosylcarboxyaminoimidazole (NCAIR) mutase, previously thought to be a
subunit of NCAIR synthetase but recently identified as an individual enzyme
[<a href="/books/n/sef/A727/?report=reader#A1295">567</a>,<a href="/books/n/sef/A727/?report=reader#A1311">583</a>], has the typical phyletic pattern of purine
biosynthesis enzymes, identical to the phyletic patterns PurF, PurD, and
PurL.</p></div><div id="A440"><h3>Phosphoribosylaminoimidazolesuccinocarboxamide synthase (EC 4.3.3.2)</h3><p>Phosphoribosylaminoimidazolesuccinocarboxamide (SAICAR) synthase (PurC) contains
a distinct version of the ATP-grasp domain. In addition to the standard set of
organisms that are capable of purine biosynthesis, SAICAR synthase is encoded in
the genome of <i>R. prowazekii</i>. It is hard to imagine what might
be the function of this enzyme in an intracellular parasite, which lacks all
other enzymes of purine biosynthesis. The sequence of <i>R.
prowazekii</i> PurC is closely related to the enzymes from other
&#x003b1;-proteobacteria but has at least three substitutions of amino acid
residues that are otherwise conserved in SAICAR synthases (E.V.K., unpublished
observations). This suggests that rickettsial SAICAR synthase might have lost
its enzymatic activity and acquired another, perhaps regulatory function.</p></div><div id="A441"><h3>Adenylosuccinate lyase (EC 4.3.2.2)</h3><p>Adenylosuccinate lyase (PurB) has the typical phyletic pattern of purine
biosynthesis enzymes, with the addition of <i>H. pylori</i>. This is
most likely due to the involvement of PurB in the conversion of IMP into AMP,
the reaction that appears to occur in <i>H. pylori</i>.</p></div><div id="A442"><h3>AICAR transformylase (EC 2.1.2.3)</h3><p>Phosphoribosylaminoimidazolecarboxamide (AICAR) formyltransferase (PurH)
catalyzes the transfer of the formyl group from formyltetrahydrofolate to AICAR.
In every organism studied to date, this protein is fused to the IMP
cyclohydrolase in a bifunctional enzyme. AICAR transformylase comprises the
C-terminal 300-aa portion of the PurH protein, whereas IMP cyclohydrolase
comprises the N-terminal 200-aa region [<a href="/books/n/sef/A727/?report=reader#A1421">693</a>]. AICAR transformylase is encoded in almost the same set of
organisms as all other purine biosynthesis enzymes, with the exception of
<i>A. fulgidus</i>, which encodes only the IMP cyclohydrolase
portion of PurH, and methanogens and pyrococci that do not encode either of
these enzymes.</p></div><div id="A443"><h3>IMP cyclohydrolase (EC 3.5.4.10)</h3><p>IMP cyclohydrolase, which catalyzes the last step of purine biosynthesis, is
fused to AICAR transformylase in every organism, except for <i>A.
fulgidus</i>, which does not have an AICAR transformylase at all, and
<i>Halobacterium</i> sp., in which the AICAR transformylase domain
is fused to PurN, a different folate-dependent GAR transformylase (see above).
As noted above, methanogens and pyrococci do not encode a recognizable IMP
cyclohydrolase.</p></div><div id="A444"><h3>Adenylosuccinate synthase (EC 6.3.4.4)</h3><p>Conversion of IMP into AMP can occur in one step, which is catalyzed by the
eukaryote-specific enzyme AMP deaminase (EC 3.5.4.6), or in two steps, as in
most bacteria and archaea. First, IMP is converted into adenylosuccinate by
adenylosuccinate synthase PurA. In addition to the entire set of organisms that
encode enzymes of IMP biosynthesis, PurA is also encoded in <i>H.
pylori</i>. The second step, the conversion of adenylosuccinate into
AMP, is catalyzed by adenylosuccinate lyase PurB (see above), which has the same
phyletic pattern as PurA.</p></div><div id="A445"><h3>IMP dehydrogenase (EC 1.1.1.205)</h3><p>Although the reverse reaction, catalyzed by GMP reductase (EC 1.6.6.8), occurs in
one step, conversion of IMP into GMP takes two steps. First, IMP is oxidized
into XMP by IMP dehydrogenase GuaB, a close paralog of GMP reductase, which,
however, contains an ~120-amino acid insert comprising two CBS domains involved
in allosteric regulation of the enzyme activity [<a href="/books/n/sef/A727/?report=reader#A807">79</a>,<a href="/books/n/sef/A727/?report=reader#A1669">941</a>]. Because
CBS is a &#x0201c;promiscuous&#x0201d; domain, which is found in association
with various proteins [<a href="/books/n/sef/A727/?report=reader#A754">26</a>], it has
caused numerous errors in automated genome annotation (see <a href="/books/n/sef/A264/?report=reader#A278">5.2.2</a>) [<a href="/books/n/sef/A727/?report=reader#A992">264</a>].
Thus, at least twelve <i>A. fulgidus</i> proteins have been annotated
as IMP dehydrogenases or &#x0201c;IMP dehydrogenase-related&#x0201d;
proteins [<a href="/books/n/sef/A727/?report=reader#A1172">444</a>], whereas, ironically,
the real IMP dehydrogenase appears not to be encoded in the <i>A.
fulgidus</i> genome. With the exception of this archaeon, IMP
dehydrogenase is present in almost every bacterial and archaeal genome sequenced
to date, including <i>A. pernix</i>, <i>C. pneumoniae</i>,
and <i>B. burgdorferi</i>, which do not encode any enzymes of IMP
biosynthesis and apparently have to import this nucleotide.</p></div><div id="A446"><h3>GMP synthase (EC 6.3.5.2)</h3><p>GMP synthase is another amidotransferase that consists of two subunits, a
glutamine amidotransferase of Triad family [<a href="/books/n/sef/A727/?report=reader#A1664">936</a>] and a synthetase subunit, which belongs to the PP-loop
superfamily of ATP pyrophosphatases [<a href="/books/n/sef/A727/?report=reader#A830">102</a>]. The phylogenetic pattern of both GMP synthase subunits is the same
as that of IMP dehydrogenase, with the addition of <i>A. fulgidus</i>,
i.e. this enzyme is also found in <i>A. pernix</i>, <i>C.
pneumoniae</i>, and <i>B. burgdorferi</i>, which lack many
other purine biosynthesis enzymes. In bacteria, yeast, and other eukaryotes, and
in <i>A. pernix</i>, these two subunits are fused together in the same
polypeptide, whereas in other archaea, they are encoded by separate genes.</p><div id="A447"><h4>General notes on purine biosynthesis evolution</h4><p>The phyletic distribution of purine biosynthesis enzymes shows some of the
trends noted above for other pathways, i.e. non-orthologous gene
displacement and increased loss of enzymes for upstream steps of the pathway
as compared to the downstream steps. With the exception of several obligate
parasites with very small genomes and <i>Buchnera</i> sp., most
bacteria encode the entire set of purine biosynthesis enzymes; there is
little doubt that they are all capable of IMP formation. Based on their gene
content, bacteria <i>H. pylori</i>, <i>C.
pneumoniae</i>, and <i>B. burgdorferi</i> and the archaeon
<i>A. pernix</i> are only capable of converting IMP into GMP;
AMP formation in these organisms probably occurs through the activity of
adenine phosphoribosyltransferase or some other mechanism. While
<i>Halobacterium</i> sp. and <i>Thermoplasma</i>
spp. encode all the enzymes of purine biosynthesis, other archaea appear to
miss at least two <i>pur</i> genes. Methanogens and pyrococci lack
<i>purK</i> and <i>purH</i> genes, and <i>A.
fulgidus</i> additionally lacks
<i>purN</i>/<i>purT</i> and <i>guaB</i>,
making it hard to judge whether the purine biosynthesis pathway is
functional in this organism. Purine biosynthesis is much more likely to
occur in methanogens and pyrococci, which would then need to harbor
alternative versions of AICAR transformylase and IMP cyclohydrolase and,
potentially, an alternative version of AIR carboxylase. Thus far, no obvious
candidates for these activities have been identified by comparative genome
analysis of these organisms. It is amazing that, although purine
biosynthesis has been intensely studied for over 50 years, comparative
genomics reveals unsuspected gaps in our understanding of this pathway and
may eventually lead to the discovery of novel enzymes.</p></div></div></div><div id="A448"><h2 id="_A448_">7.4. Amino Acid Biosynthesis</h2><div id="A449"><h3>7.4.1. Aromatic amino acids</h3><div id="A450"><h4>7.4.1.1. Common steps of the pathway</h4><p>The biosynthetic pathways for phenylalanine, tyrosine, and tryptophan in
bacteria and eukaryotes share common steps leading from phosphoenolpyruvate
and erythrose-4-phosphate to chorismate. Enzymes for most of these steps are
encoded also in archaeal genomes.</p><div id="A451"><h5>2-Dehydro-3-deoxy-D-arabino-heptonate 7-phosphate synthase (EC
4.1.2.15)</h5><p>Although 2-dehydro-3-deoxy-D-arabino-heptonate 7-phosphate (DAHP)
synthase is found in <i>E. coli</i> in three different
versions, AroF, AroG, and AroH, all of these enzymes are close paralogs
and represent the so-called microbial form of DAHP synthase. A different
form of this enzyme was originally described in potato and
<i>Arabidopsis</i> and designated the plant form.
Subsequently, this form has been discovered also in bacteria [<a href="/books/n/sef/A727/?report=reader#A933">205</a>,<a href="/books/n/sef/A727/?report=reader#A1026">298</a>,<a href="/books/n/sef/A727/?report=reader#A1161">433</a>,<a href="/books/n/sef/A727/?report=reader#A1607">879</a>]. This form
is encoded in many complete genomes and is the only DAHP synthase in
<i>M. tuberculosis</i>, <i>M. leprae</i>,
<i>H. pylori</i>, and <i>C. jejuni</i> (<a class="figpopup" href="/books/NBK20266/figure/A452/?report=objectonly" target="object" rid-figpopup="figA452" rid-ob="figobA452">Figure 7.6</a>). <i>B.
subtilis</i> and several other Gram-positive bacteria encode a
third form of DAHP synthase, referred to as AroA(G), which is homologous
to 3-deoxy-D-manno-octulosonate 8-phosphate synthase of <i>E.
coli</i> [<a href="/books/n/sef/A727/?report=reader#A824">96</a>].
Remarkably, this third form is also found in <i>T.
maritima</i> and in several archaea, such as <i>P.
abyssi</i>, <i>A. pernix</i>, and
<i>Thermoplasma</i> spp. Other archaea, such as <i>A.
fulgidus</i>, <i>M. jannaschii</i>, and <i>M.
thermoautotrophicum</i>, as well as the bacterium <i>A.
aeolicus</i>, encode neither of these three DAHP synthases and
appear to synthesize 3-dehydroquinate via a different mechanism that
does not include DAHP as an intermediate (see below).
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA452" co-legend-rid="figlgndA452"><a href="/books/NBK20266/figure/A452/?report=objectonly" target="object" title="Figure 7.6" class="img_link icnblk_img figpopup" rid-figpopup="figA452" rid-ob="figobA452"><img class="small-thumb" src="/books/NBK20266/bin/ch7f6.gif" src-large="/books/NBK20266/bin/ch7f6.jpg" alt="Figure 7.6. Distribution of tryptophan biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA452"><h4 id="A452"><a href="/books/NBK20266/figure/A452/?report=objectonly" target="object" rid-ob="figobA452">Figure 7.6</a></h4><p class="float-caption no_bottom_margin">Distribution of tryptophan biosynthesis enzymes in
organisms with completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A453"><h5>3-Dehydroquinate synthase (EC 4.6.1.3)</h5><p>3-Dehydroquinate synthase is found in many bacteria and in some archaea.
Remarkably, the phyletic pattern of this enzyme exactly corresponds to
the overlap of the phyletic patterns for the three forms of DAHP
synthase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e9.jpg" alt="Image ch7e9.jpg" /></div><p>This correlation suggests that a single form of 3-dehydroquinate synthase
can account for the conversion of DAHP into 3-dehydroquinate in all the
organisms with completely sequenced genomes and probably represents the
only form of this enzyme.</p></div><div id="A454"><h5>3-Dehydroquinate dehydratase (EC 4.2.1.10)</h5><p>Two forms of 3-dehydroquinate dehydratase have been characterized and
designated class I (encoded by <i>aroD</i> gene) and class II
(encoded by <i>aroQ</i> or QUTE genes), respectively. Taken
together, these two enzymes completely cover the phyletic diversity of
the organisms that encode 3-dehydroquinate synthase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e10.jpg" alt="Image ch7e10.jpg" /></div><p>Notably, dehydroquinate dehydratase (as well as most of the other enzymes
of tryptophan biosynthesis) is found in several genomes that do not
encode dehydroquinate synthase, indicating the existence of an
alternative, still uncharacterized pathway of dehydroquinate formation
in <i>Halobacterium</i> sp., <i>A. fulgidus</i>,
<i>M. jannaschii</i>, and <i>M.
thermoautotrophicum</i>, and <i>A. aeolicus.</i>
</p></div><div id="A455"><h5>Shikimate 5-dehydrogenase (EC 1.1.1.25)</h5><p>The last enzyme in the shikimate-producing part of the pathway, shikimate
5-dehydrogenase, is encoded in most of the completely sequenced
bacterial (with the exception of rickettsiae, spirochetes, and
mycoplasmas) and archaeal (with the exception of <i>P.
horikoshii</i>) genomes. The phyletic pattern for shikimate
5-dehydrogenase coincides with the combined pattern of the two forms of
3-dehydroquinate synthase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e11.jpg" alt="Image ch7e11.jpg" /></div></div><div id="A456"><h5>Shikimate kinase (EC 2.7.1.71)</h5><p>The typical form of shikimate kinase, found in bacteria and eukaryotes,
is not encoded in any archaeal genome sequenced so far. Recently, a
shikimate kinase of the GHMP superfamily has been identified and
experimentally studied in <i>M. jannaschii</i> [<a href="/books/n/sef/A727/?report=reader#A899">171</a>]. This enzyme is encoded in
each archaeal genome, except for <i>P. horikoshii</i>.
Together, these two forms of shikimate kinase have the same phyletic
pattern as the combination of the two forms of 3-dehydroquinate
dehydratase or shikimate dehydrogenase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e12.jpg" alt="Image ch7e12.jpg" /></div></div><div id="A457"><h5>5-Enolpyruvylshikimate 3-phosphate synthase (EC 2.5.1.19)</h5><p>Like shikimate dehydrogenase, 5-enolpyruvylshikimate 3-phosphate synthase
(AroA) is found in only one form with the same phyletic pattern as the
preceding enzyme.</p></div><div id="A458"><h5>Chorismate synthase (EC 4.6.1.4)</h5><p>The only known chorismate synthase (AroC) has the same phyletic pattern
as shikimate dehydrogenase and 5-enolpyruvylshikimate 3-phosphate
synthase.</p></div></div><div id="A459"><h4>7.4.1.2. Tryptophan biosynthesis</h4><p>After chorismate, the tryptophan biosynthetic pathway deviates from the
pathways leading to phenylalanine and tyrosine. In the tryptophan branch,
all the remaining enzymes have very similar phyletic patterns.</p><div id="A460"><h5>Anthranilate synthase (EC 4.1.3.27)</h5><p>Anthranilate synthase and the closely related para-aminobenzoate synthase
consist of two components, the synthetase subunit and the glutamine
amidotransferase subunit, which, in most organisms, are encoded by
separate genes <i>trpG</i> (or <i>pabA</i>) and
<i>trpE</i> (or <i>pabB</i>). In <i>E.
coli</i>, the <i>trpG</i> gene for glutamine
amidotransferase subunit is fused to the <i>trpD</i> gene that
encodes anthranilate phosphoribosyltransferase, the enzyme catalyzing
the next step of the pathway. This sometimes leads to a confusion in
nomenclature, with the <i>trpG</i> gene being referred to as
<i>trpD</i> or as
<i>trpD</i>_<i>1</i>. The phyletic pattern for
anthranilate synthase is the same as the patterns described above for
shikimate dehydrogenase, 5-enolpyruvylshikimate 3-phosphate synthase,
and chorismate synthase, with the exception that anthranilate synthase
is missing in chlamydiae.</p></div><div id="A461"><h5>Anthranilate phosphoribosyltransferase (EC 2.4.2.18)</h5><p>There is only one form of anthranilate phosphoribosyltransferase that
shows almost the same phyletic pattern as anthranilate synthase. The
only diversification in the existing set of genomes is the absence of
the <i>trpD</i> gene (as well as genes for the remaining steps
of tryptophan biosynthesis) in <i>S. pyogenes</i>. This
probably means that genes annotated as <i>trpG</i> and
<i>trpE</i> in <i>S. pyogenes</i> actually
encode para-aminobenzoate synthase and have no role in tryptophan
biosynthesis.</p></div><div id="A462"><h5>N-(5&#x02032;-Phosphoribosyl)anthranilate isomerase (EC
5.3.1.24)</h5><p>Phosphoribosylanthranilate isomerase is also represented by only one form
with essentially the same phyletic pattern as anthranilate
phosphoribosyltransferase. Here again, the nomenclature is somewhat
complicated because of a gene fusion in <i>E. coli</i>. The
phosphoribosylanthranilate isomerase gene that, in most species, is
referred to as <i>trpF,</i> in <i>E. coli</i> is
fused to the <i>trpC</i> gene that encodes indole-3-glycerol
phosphate synthase, the enzyme for the next step of the pathway.
Therefore, in <i>E. coli,</i> the <i>trpF</i> gene
is sometimes also referred to as <i>trpC</i>, which can lead
to confusion. The phyletic pattern of phosphoribosylanthranilate
isomerase is essentially the same as that of other enzymes of tryptophan
biosynthesis, with the most notable difference being the apparent
absence of <i>trpF</i> in <i>M. tuberculosis</i> and
<i>M. leprae</i>. Another peculiarity is the unusual
distribution of phosphoribosylanthranilate isomerase in different
chlamydial species: while <i>C. trachomatis</i> and <i>C.
muridarum</i> both have the <i>trpF</i> gene,
<i>C. pneumoniae</i> does not. This probably reflects the
ongoing gene loss in the evolution of chlamydiae.</p></div><div id="A463"><h5>Indole-3-glycerol phosphate synthase (EC 4.1.1.48)</h5><p>Indole-3-glycerol phosphate synthase exists in a single form with the
same phyletic pattern as anthranilate phosphoribosyltransferase.</p></div><div id="A464"><h5>Tryptophan synthase (EC 4.2.1.20)</h5><p>Tryptophan synthase consists of two subunits, which are encoded by
<i>trpA</i> and <i>trpB</i> genes. Their
phyletic patterns are similar to that of anthranilate
phosphoribosyltransferase, with the exception that, as seen above for
<i>trpC</i>, <i>trpA</i>, and
<i>trpB</i> genes are found in <i>C.
trachomatis</i> and <i>C. muridarum</i> but not in
<i>C. pneumoniae</i>. </p></div></div><div id="A465"><h4>7.4.1.3. Phenylalanine and tyrosine biosynthesis</h4><div id="A466"><h5>Chorismate mutase (EC 5.4.99.5)</h5><p>Chorismate mutase is involved in both phenylalanine and tyrosine
biosynthesis. The best-known version of this enzyme is found in
<i>E. coli</i> in two paralogous forms fused with
prephenate dehydratase in PheA and prephenate dehydrogenase in TyrA. In
addition to these two forms, there is (i) a distantly related form of
chorismate mutase encoded in yeast, fungi, and in plant cells and (ii)
an unrelated monofunctional form found in <i>B. subtilis</i>,
<i>Synechocystis</i> sp., and many other Gram-positive
bacteria and cyanobacteria.</p><p>Although these three forms of chorismate mutase show almost no sequence
similarity to each other, structural comparisons indicate that the
<i>E. coli</i> and yeast enzymes are related to each other
and are unrelated to the form found in <i>B. subtilis</i> and
<i>Th. thermophilus</i>. A comparison of the combined
phyletic pattern of all these forms of chorismate mutase with that of
chorismate synthase shows that, with the exception of <i>P.
abyssi</i> and in <i>Chlamydia</i> spp., all
organisms that produce chorismate are capable of converting it to
prephenate.</p></div><div id="A467"><h5>Prephenate dehydrogenase (EC 1.3.1.12)</h5><p>Prephenate dehydrogenase TyrA, an enzyme of the tyrosine biosynthesis
branch of the pathway, is found in a single form with almost the same
phyletic pattern as chorismate mutase (<a class="figpopup" href="/books/NBK20266/figure/A468/?report=objectonly" target="object" rid-figpopup="figA468" rid-ob="figobA468">Figure 7.7</a>). The only exception is <i>S.
pyogenes</i> that appears not to encode prephenate
dehydrogenase.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA468" co-legend-rid="figlgndA468"><a href="/books/NBK20266/figure/A468/?report=objectonly" target="object" title="Figure 7.7" class="img_link icnblk_img figpopup" rid-figpopup="figA468" rid-ob="figobA468"><img class="small-thumb" src="/books/NBK20266/bin/ch7f7.gif" src-large="/books/NBK20266/bin/ch7f7.jpg" alt="Figure 7.7. Distribution of phenylalanine and tyrosine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA468"><h4 id="A468"><a href="/books/NBK20266/figure/A468/?report=objectonly" target="object" rid-ob="figobA468">Figure 7.7</a></h4><p class="float-caption no_bottom_margin">Distribution of phenylalanine and tyrosine biosynthesis
enzymes in organisms with completely sequenced
genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A469"><h5>Prephenate dehydratase (EC 4.2.1.51)</h5><p>Prephenate dehydratase, an enzyme of the phenylalanine biosynthesis
branch of the pathway, is also represented by a single form in all known
organisms. However, its phyletic pattern shows the absence of this
enzyme in <i>S. pyogenes</i>, <i>H. pylori</i>, and
<i>A. pernix</i>, which are all capable of producing
prephenate, suggesting that these organisms either lack phenylalanine
biosynthesis or have an alternative form of prephenate dehydratase.</p></div><div id="A470"><h5>Aromatic aminotransferase (EC 2.6.1.1, 2.6.1.5, 2.6.1.9,
2.6.1.57)</h5><p>There are several families of pyridoxal-phosphate-dependent
aminotransferases that are capable of producing tyrosine and
phenylalanine from, respectively, 4-hydroxyphenylpyruvate and
phenylpyruvate. Although the best-studied tyrosine aminotransferase,
<i>E. coli</i> TyrB, has a relatively narrow phyletic
distribution, homologs of histidinol phosphate aminotransferase and
aspartate aminotransferase are encoded in every bacterial and archaeal
genome except for spirochetes and mycoplasmas. Thus, once phenylpyruvate
and 4-hydroxyphenylpyruvate are synthesized, their transamination into,
respectively, phenylalanine and tyrosine can be performed by all
organisms whose genome sequences are currently available.</p><div id="A471"><h5>A summary on aromatic amino acid biosynthesis</h5><p>Because aromatic amino acid biosynthesis shares common steps with
biosynthesis of ubiquinone, this pathway displays a stunning variety
of alternative enzymes catalyzing the same reaction. This makes
analysis of their phyletic patterns rather complicated, but, at the
same time, allows one to draw some interesting conclusions. Most
bacteria and archaea retain the complete set of genes for tryptophan
biosynthesis. The exceptions are the obligate archaeal heterotroph
<i>P. horikoshii</i> and some obligate bacterial
parasites, such as <i>S. pyogenes</i>, rickettsiae,
chlamydiae, spirochetes, and mycoplasmas, which apparently obtain
tryptophan, just like many other nutrients, from other microbes and
from the host, respectively.</p><p>Enzymes of the tyrosine biosynthesis pathway are encoded in almost as
many complete genomes, with the conspicuous exception of <i>P.
abyssi</i>. One could speculate that, while tryptophan is
rapidly degraded at 105&#x000ba;C (the optimal growth temperature
of this organism), tyrosine is not, which alleviates the requirement
for <i>de novo</i> synthesis. These considerations could
also explain the absence of phenylalanine biosynthesis in <i>P.
abyssi</i> and <i>A. pernix</i>.</p><p>The consistency of the phyletic patterns of the enzymes for the
downstream stages of aromatic amino acid biosynthesis underscores
the remaining problem with the early stages. Indeed, <i>A.
aeolicus</i> and four archaeal species encode
3-dehydroquinate dehydratase and all the downstream enzymes but do
not encode either DAHP synthase or 3-dehydroquinate synthase:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e13.jpg" alt="Image ch7e13.jpg" /></div><p>It appears that these organisms produce 3-dehydroquinate via a
different mechanism, which does not include DAHP as an intermediate.
Using the COG phyletic pattern search tool, one could search for
orthologous protein sets that are represented in those five genomes
but are missing in thermoplasmas, pyrococci, <i>A.
pernix</i>, and <i>T. maritima</i>, all of which
encode a DAHP synthase and a 3-dehydroquinate synthase. Such a
search identified just four COGs, only one of which, COG1465,
consisted of uncharacterized proteins. These proteins, orthologs of
<i>M. jannaschii</i> MJ1249, can be predicted to
function as an alternative 3-dehydroquinate synthases (M.Y.G.,
unpublished). This prediction seems to be further supported by the
adjacency of the genes encoding COG1465 members AF0229 and VNG0310C
to the <i>aroC</i> gene in the genomes of <i>A.
fulgidus</i> and <i>Halobacterium</i> sp.,
respectively. However, even if this prediction is correct, the exact
nature of the precursor for 3-dehydroquinate and the mechanism of
its biosynthesis in these organisms need to be elucidated
experimentally.</p></div></div></div></div><div id="A472"><h3>7.4.2. Arginine biosynthesis</h3><div id="A473"><h4>N-Acetylglutamate synthase (EC 2.3.1.1, 2.3.1.35)</h4><p>The first step in arginine biosynthesis from glutamate is its acetylation,
with either acetyl-CoA or acetylornithine utilized as donors of the acetyl
group (<a class="figpopup" href="/books/NBK20266/figure/A474/?report=objectonly" target="object" rid-figpopup="figA474" rid-ob="figobA474">Figure 7.8</a>). In <i>E.
coli</i> and several other organisms, this reaction is catalyzed by
the acetyltransferase ArgA, which employs acetyl-CoA as the acetyl donor. In
all proteobacteria that encode this enzyme, the <i>argA</i> gene
is fused to the gene for N-acetylglutamate kinase, which catalyzes the next
step of the pathway. Like in other domain fusion cases, confusion
occasionally emerges during genome annotation, especially because the
N-terminal kinase domain, which consists of ~300 amino acid residues, can
make the C-terminal acetyltransferase domain almost invisible in BLAST
outputs (see <a href="/books/n/sef/A166/?report=reader#A224">4.4.4</a>).
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA474" co-legend-rid="figlgndA474"><a href="/books/NBK20266/figure/A474/?report=objectonly" target="object" title="Figure 7.8" class="img_link icnblk_img figpopup" rid-figpopup="figA474" rid-ob="figobA474"><img class="small-thumb" src="/books/NBK20266/bin/ch7f8.gif" src-large="/books/NBK20266/bin/ch7f8.jpg" alt="Figure 7.8. Distribution of arginine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA474"><h4 id="A474"><a href="/books/NBK20266/figure/A474/?report=objectonly" target="object" rid-ob="figobA474">Figure 7.8</a></h4><p class="float-caption no_bottom_margin">Distribution of arginine biosynthesis enzymes in organisms
with completely sequenced genomes. All details as in Figure
2.7. </p></div></div><p>A different, unrelated N-acetylglutamate synthase (N-acetylornithine
transferase, the <i>argJ</i> gene product) is present in
<i>B. subtilis</i>, yeast, and many other organisms. This
enzyme couples acetylation of glutamate with deacetylation of
N-acetylornithine, which is the fifth step in arginine biosynthesis. This
activity allows recycling of the acetyl group in the arginine biosynthesis
pathway.</p></div><div id="A475"><h4>N-Acetylglutamate kinase (EC 2.7.2.8)</h4><p>Phosphorylation of N-acetylglutamate is catalyzed by the product of the
<i>argB</i> gene, a kinase with the carbamate kinase fold.
This enzyme is found in a wide variety of organisms, such that its phyletic
pattern is even broader than the combined patterns of both enzymes that
generate N-acetylglutamate:</p><div class="graphic"><img src="/books/NBK20266/bin/ch7e14.jpg" alt="Image ch7e14.jpg" /></div><p>However, N-acetylglutamate kinase is not encoded in the genomes of many
parasitic bacteria, such as <i>S. pyogenes</i>, <i>H.
influenzae</i>, <i>H. pylori</i>, chlamydiae,
rickettsiae, spirochetes, and mycoplasmas.</p></div><div id="A476"><h4>N-Acetyl-gamma-glutamyl phosphate reductase (EC 1.2.1.38)</h4><p>The enzyme that catalyzes the next step of the pathway, ArgC, has the same
phyletic pattern as ArgB. In fungi, <i>argB</i> and
<i>argC</i> genes are fused and encode a single bifunctional
protein.</p></div><div id="A477"><h4>N-Acetylornithine aminotranferase (EC 2.6.1.11)</h4><p>N-Acetylornithine deacetylase (N-acetylornithinase) belongs to a large family
of closely related acetyltransferases (deacetylases), which is represented
by two or more paralogs even in the relatively small genomes of <i>H.
influenzae</i>, <i>L. lactis</i>, and <i>S.
pyogenes</i>. Although proper assignment of substrate specificity
in such a case is difficult, if not impossible, the few organisms that
produce N-acetylornithine but lack the ArgJ-type N-acetylglutamate synthase
offer an ample choice of candidates for the function of
N-acetylornithinase.</p></div><div id="A478"><h4>N-Acetylornithine deacetylase (EC 3.5.1.16)</h4><p>N-Acetylornithinase belongs to a large family of closely related
acetyltransferases (deacylases), which is represented by two or more
paralogs even in the relatively small genomes of <i>H.
influenzae</i>, <i>L. lactis</i>, and <i>S.
pyogenes</i>. Although proper assignment of substrate specificity
in such a case is difficult, if not impossible, the few organisms that
produce N-acetylornithine but lack the ArgJ-type N-acetylglutamate synthase
offer an ample choice of candidates for the role of N-acetylornithinase.</p></div><div id="A479"><h4>Ornithine carbamoyltransferase (EC 2.1.3.6)</h4><p>Ornithine carbamoyltransferase catalyzes the sixth step of arginine
biosynthesis, conversion of ornithine into citrulline. Carbamoyl phosphate
that serves as the second substrate of this reaction is provided by
carbamoyl phosphate synthetase, which was discussed above (see <a href="#A419">7.2</a>). Ornithine carbamoyltransferase
has a much wider phyletic distribution than other enzymes of arginine
biosynthesis. This is probably due to the fact that it also catalyzes the
reverse reaction, i. e. phosphorolysis of citrulline with the formation of
ornithine and carbamoyl phosphate, which is part of the urea cycle.
Accordingly, ornithine carbamoyltransferase is found in humans and other
higher eukaryotes, which have the urea cycle but are incapable of arginine
biosynthesis.</p></div><div id="A480"><h4>Argininosuccinate synthase (EC 6.3.4.5)</h4><p>Like ornithine carbamoyltransferase, argininosuccinate synthase participates
in the urea cycle. As a result, its phyletic distribution is also wider than
that of the enzymes that catalyze early steps of arginine biosynthesis. This
enzyme, too, is found in humans and in bacteria, such as <i>H.
influenzae</i>, which have all the urea cycle enzymes but lack
several enzymes of arginine biosynthesis.</p></div><div id="A481"><h4>Argininosuccinate lyase (EC 4.3.2.1)</h4><p>Argininosuccinate lyase, the last enzyme of arginine biosynthesis, splits
argininosuccinate into arginine and fumarate. Like the two preceding
enzymes, it also participates in the urea cycle, and its phyletic pattern is
nearly identical to that of argininosuccinate synthetase.</p></div></div><div id="A482"><h3>7.4.3. Histidine biosynthesis</h3><p>In contrast to the pathways of aromatic amino acid and arginine biosynthesis,
histidine biosynthesis exhibits remarkable consistency of the phyletic patterns
of all the enzymes involved (<a class="figpopup" href="/books/NBK20266/figure/A483/?report=objectonly" target="object" rid-figpopup="figA483" rid-ob="figobA483">Figure 7.9</a>).
While the first enzyme of the pathway, phosphoribosylpyrophosphate synthetase
(EC 2.7.6.1), also participates in the purine biosynthesis pathway (see above),
nearly all the committed enzymes of histidine biosynthesis have the same
phyletic pattern, indicating that this pathway is encoded in the great majority
of complete prokaryotic genomes sequenced to date. The exceptions are the
heterotrophic archaea <i>Thermoplasma</i> spp.,
<i>Pyrococcus</i> sp., and <i>A. pernix</i>, and
parasitic bacteria with small genomes, namely rickettsiae, chlamydiae,
spirochetes, and mycoplasmas, as well as <i>S. pyogenes</i> and
<i>H. pylori</i> (despite their larger genomes). Remarkably, the
aphid symbiont <i>Buchnera</i> sp., which has the second smallest
genome available to date, encodes the complete set of histidine biosynthesis
enzymes.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA483" co-legend-rid="figlgndA483"><a href="/books/NBK20266/figure/A483/?report=objectonly" target="object" title="Figure 7.9" class="img_link icnblk_img figpopup" rid-figpopup="figA483" rid-ob="figobA483"><img class="small-thumb" src="/books/NBK20266/bin/ch7f9.gif" src-large="/books/NBK20266/bin/ch7f9.jpg" alt="Figure 7.9. Distribution of histidine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA483"><h4 id="A483"><a href="/books/NBK20266/figure/A483/?report=objectonly" target="object" rid-ob="figobA483">Figure 7.9</a></h4><p class="float-caption no_bottom_margin">Distribution of histidine biosynthesis enzymes in organisms with
completely sequenced genomes. All details as in Figure
2.7. </p></div></div><p>There are several deviations from this common pattern. First, phosphoribosyl-ATP
pyrophosphatase (EC 3.6.1.31) was not detected in <i>A. fulgidus</i>.
Since this organism encodes genes for all other enzymes of histidine
biosynthesis, one should assume that this reaction in <i>A.
fulgidus</i> is catalyzed by an unrelated pyrophosphatase. Indeed,
<i>A. fulgidus</i> genome encodes several predicted
pyrophosphatases of unknown specificity (COG1694) that could be good candidates
for the role of the missing phosphoribosyl-ATP pyrophosphatase.</p><p>Another deviation from the common pattern is the existence of at least two
unrelated histidinol phosphatases (<a class="figpopup" href="/books/NBK20266/figure/A483/?report=objectonly" target="object" rid-figpopup="figA483" rid-ob="figobA483">Figure
7.9</a>), one of which has been experimentally characterized in
<i>E. coli</i> and the other in yeast and <i>B.
subtilis</i> [<a href="/books/n/sef/A727/?report=reader#A743">15</a>,<a href="/books/n/sef/A727/?report=reader#A1228">500</a>]. The latter form of this enzyme
(COG1387) belongs to a large superfamily of PHP-type phosphohydrolases [<a href="/books/n/sef/A727/?report=reader#A769">41</a>], which have common sequence motifs but
clearly differ in substrate specificity. A closer inspection of COG1387 shows
that proteins from yeast, <i>B. subtilis</i>, <i>B.
halodurans</i>, <i>L. lactis</i>, <i>D.
radiodurans</i>, and <i>T. maritima</i>, comprise a tight
orthologous set and can be confidently predicted to possess histidinol
phosphatase activity. Other members of this COG are more distantly related to
the experimentally characterized histidinol phosphatases from yeast and
<i>B. subtilis</i> and might have other substrates. In addition,
both forms of histidinol phosphatase are missing in
<i>Halobacterium</i> sp. Therefore it appears likely that there is
yet another, so far unrecognized, form of histidinol phosphatase in
<i>Halobacterium</i> sp., <i>Thermoplasma</i> spp., and
other organisms. There are plenty of unassigned predicted hydrolases that could
potentially have this activity.</p><p>Remarkably, the ortholog of the <i>E. coli</i> histidinol phosphatase
(HisB, COG0241) is encoded in <i>H. pylori</i>, which lacks all the
other enzymes of histidine biosynthesis. This protein most likely represents a
case of enzyme recruitment and functions as a phosphatase that hydrolyzes some
other phosphoester.</p></div><div id="A484"><h3>7.4.4. Biosynthesis of branched-chain amino acids</h3><p>To those readers who are already tired of numerous instances of non-orthologous
gene displacement in metabolic pathways, biosynthesis of leucine, isoleucine,
and valine offers a well-deserved reprieve. In these pathways, the only instance
of alternative enzymes catalyzing the same reaction is the last step, amination
of &#x003b1;-ketomethylvaleriate, &#x003b1;-ketoisovaleriate, and
&#x003b1;-ketoisocaproate. In addition to the branched-chain amino acid
aminotransferase IlvE, this reaction can be catalyzed by alternative
aminotransferases (<a class="figpopup" href="/books/NBK20266/figure/A485/?report=objectonly" target="object" rid-figpopup="figA485" rid-ob="figobA485">Figure 7.10</a>)
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA485" co-legend-rid="figlgndA485"><a href="/books/NBK20266/figure/A485/?report=objectonly" target="object" title="Figure 7.10" class="img_link icnblk_img figpopup" rid-figpopup="figA485" rid-ob="figobA485"><img class="small-thumb" src="/books/NBK20266/bin/ch7f10.gif" src-large="/books/NBK20266/bin/ch7f10.jpg" alt="Figure 7.10. Distribution of isoleucine/leucine/valine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA485"><h4 id="A485"><a href="/books/NBK20266/figure/A485/?report=objectonly" target="object" rid-ob="figobA485">Figure 7.10</a></h4><p class="float-caption no_bottom_margin">Distribution of isoleucine/leucine/valine biosynthesis enzymes in
organisms with completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A486"><h3>7.4.5. Proline biosynthesis</h3><p>The best characterized pathway of proline biosynthesis is a three-step chain of
reactions (<a class="figpopup" href="/books/NBK20266/figure/A487/?report=objectonly" target="object" rid-figpopup="figA487" rid-ob="figobA487">Figure 7.11</a>) that converts
glutamate into proline through consecutive action of glutamate kinase (ProB, EC
2.7.2.11), &#x003b3;-glutamyl phosphate reductase (ProA, EC 1.2.1.41), and
&#x00394;-pyrroline-5-carboxylate reductase (ProC, EC 1.5.1.2). This pathway
is encoded in yeast, <i>E. coli</i>, <i>B. subtilis</i>, and
in many other bacteria, including <i>C. jejuni</i> (but not <i>H.
pylori</i>) and <i>T. pallidum</i> (but not <i>B.
burgdorferi</i>). This pathway, however, is not detectable in archaea,
except for the two species of <i>Methanosarcina</i>, which, in all
likelihood, acquired it through HGT. Instead, <i>Halobacterium</i>
sp., <i>A. fulgidus</i>, <i>M. thermoautotrophicum</i>,
<i>Thermoplasma</i> spp., and <i>A. pernix</i> encode an
unusual enzyme, ornithine cyclodeaminase (EC 4.3.1.12), which directly makes
proline from ornithine. This enzyme, first discovered in tumor-inducing (Ti)
plasmids of <i>A. tumefaciens</i>, was later found in pseudomonads and
other bacteria [<a href="/books/n/sef/A727/?report=reader#A911">183</a>,<a href="/books/n/sef/A727/?report=reader#A1526">798</a>]. In plants, expression of this
interesting enzyme stimulates flowering [<a href="/books/n/sef/A727/?report=reader#A1578">850</a>], whereas the mammalian ortholog of this enzyme is expressed in
neural tissue, including human retina, and functions as &#x003bc;-crystallin, a
major component of the eye lens in marsupials [<a href="/books/n/sef/A727/?report=reader#A1166">438</a>]. Although no such gene was detected in <i>M.
jannaschii</i>, this archaeon, too, has been reported to possess
ornithine cyclodeaminase activity [<a href="/books/n/sef/A727/?report=reader#A1037">309</a>].
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA487" co-legend-rid="figlgndA487"><a href="/books/NBK20266/figure/A487/?report=objectonly" target="object" title="Figure 7.11" class="img_link icnblk_img figpopup" rid-figpopup="figA487" rid-ob="figobA487"><img class="small-thumb" src="/books/NBK20266/bin/ch7f11.gif" src-large="/books/NBK20266/bin/ch7f11.jpg" alt="Figure 7.11. Distribution of proline biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA487"><h4 id="A487"><a href="/books/NBK20266/figure/A487/?report=objectonly" target="object" rid-ob="figobA487">Figure 7.11</a></h4><p class="float-caption no_bottom_margin">Distribution of proline biosynthesis enzymes in organisms with
completely sequenced genomes. </p></div></div><p>An interesting aspect of proline metabolism is that its biosynthesis and
degradation both proceed through the &#x00394;-pyrroline-5-carboxylate
intermediate. As a result, the proline biosynthetic pathway is sometimes
confused with proline catabolism. Another complication in the analysis of
proline metabolism is that, in <i>E. coli</i> and several other
bacteria, the genes for proline dehydrogenase (EC 1.5.99.8) and
&#x003b3;-glutamate semialdehyde dehydrogenase (EC 1.5.1.12), the first and
second enzymes of proline catabolism, respectively, are fused, forming the
bifunctional protein PutA. In the COG database, these two domains of the PutA
protein belong to two different COGs, COG0506 and COG1012 (<a class="figpopup" href="/books/NBK20266/figure/A487/?report=objectonly" target="object" rid-figpopup="figA487" rid-ob="figobA487">Figure 7.11</a>).</p><p>In conclusion, proline metabolism is tightly interlinked with arginine
metabolism. Proline biosynthesis from glutamate can be reconstructed in all
organisms with completely sequenced genomes with the exception of pyrococci,
<i>H. pylori</i>, <i>B. burgdorferi</i>, chlamydiae, and
mycoplasmas. The gene encoding ornithine cyclodeaminase in <i>M.
jannaschii</i> [<a href="/books/n/sef/A727/?report=reader#A1037">309</a>] remains
to be identified. It can be expected to be a member of a different enzyme
family, unrelated to the known ornithine cyclodeaminases (COG2423).</p></div></div><div id="A488"><h2 id="_A488_">7.5. Coenzyme Biosynthesis</h2><div id="A489"><h3>7.5.1. Thiamine</h3><p>Biosynthesis of cofactors (coenzymes), particularly thiamine, is a surprisingly
poorly studied area of biochemistry. Although the first <i>thi</i>
mutations in <i>E. coli</i> were characterized half a century ago, the
complete list of thiamine biosynthesis genes has been determined only in the
1990's [<a href="/books/n/sef/A727/?report=reader#A1596">868</a>], and the functions of
their products have been characterized only in the last several years [<a href="/books/n/sef/A727/?report=reader#A809">81</a>,<a href="/books/n/sef/A727/?report=reader#A811">83</a>,<a href="/books/n/sef/A727/?report=reader#A1655">927</a>]. The scheme for
thiamine biosynthesis in <a class="figpopup" href="/books/NBK20266/figure/A490/?report=objectonly" target="object" rid-figpopup="figA490" rid-ob="figobA490">Figure 7.12</a> was
drawn using the <i>E. coli</i> data. One cannot help noticing that
every enzyme on this chart has its own distinct phyletic pattern. This indicates
the abundance of non-orthologous gene displacement cases among thiamine
biosynthesis enzymes and suggests that different organisms might use different
compounds as thiamin precursors. The apparent absence of ThiC in thermoplasmas,
<i>A. pernix</i>, <i>H. influenzae</i>, and <i>H.
pylori</i>, all of which encode ThiD (<a class="figpopup" href="/books/NBK20266/figure/A490/?report=objectonly" target="object" rid-figpopup="figA490" rid-ob="figobA490">Figure 7.12</a>), is a strong indication that some intermediate other
than AIR is used as a precursor in these organisms. Thus, although all steps of
the thiamine biosynthesis pathway have been resolved for <i>E.
coli</i> [<a href="/books/n/sef/A727/?report=reader#A809">81</a>], there is still
ample opportunity for new discoveries in other organisms.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA490" co-legend-rid="figlgndA490"><a href="/books/NBK20266/figure/A490/?report=objectonly" target="object" title="Figure 7.12" class="img_link icnblk_img figpopup" rid-figpopup="figA490" rid-ob="figobA490"><img class="small-thumb" src="/books/NBK20266/bin/ch7f12.gif" src-large="/books/NBK20266/bin/ch7f12.jpg" alt="Figure 7.12. Distribution of thiamine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA490"><h4 id="A490"><a href="/books/NBK20266/figure/A490/?report=objectonly" target="object" rid-ob="figobA490">Figure 7.12</a></h4><p class="float-caption no_bottom_margin">Distribution of thiamine biosynthesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A491"><h3>7.5.2. Riboflavin</h3><p>The riboflavin biosynthesis pathway is a challenging case, with three of the
seven <i>rib</i> genes characterized in <i>E. coli</i> and
<i>B. subtilis</i> having no archaeal orthologs (<a class="figpopup" href="/books/NBK20266/figure/A492/?report=objectonly" target="object" rid-figpopup="figA492" rid-ob="figobA492">Figure 7.13</a>). The archaeal variant of
riboflavin synthase, the last enzyme of the pathway, has been identified and
turned out to be unrelated to the bacterial enzyme [<a href="/books/n/sef/A727/?report=reader#A935">207</a>]. In contrast, the archaeal versions of the first two
enzymes of the pathway, GTP cyclohydrolase II (RibA) and pyrimidine deaminase
(RibD1), remain unknown, so there is an excellent chance of discovering new
enzymes of this pathway.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA492" co-legend-rid="figlgndA492"><a href="/books/NBK20266/figure/A492/?report=objectonly" target="object" title="Figure 7.13" class="img_link icnblk_img figpopup" rid-figpopup="figA492" rid-ob="figobA492"><img class="small-thumb" src="/books/NBK20266/bin/ch7f13.gif" src-large="/books/NBK20266/bin/ch7f13.jpg" alt="Figure 7.13. Distribution of riboflavin biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA492"><h4 id="A492"><a href="/books/NBK20266/figure/A492/?report=objectonly" target="object" rid-ob="figobA492">Figure 7.13</a></h4><p class="float-caption no_bottom_margin">Distribution of riboflavin biosynthesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A493"><h3>7.5.3. NAD</h3><p>Nicotinate mononucleotide adenylyltransferase, the last missing enzyme of the NAD
biosynthesis pathway, has been characterized only in 2000, thanks in part to the
genome context-based methods [<a href="/books/n/sef/A727/?report=reader#A810">82</a>,<a href="/books/n/sef/A727/?report=reader#A1291">563</a>,<a href="/books/n/sef/A727/?report=reader#A1320">592</a>]. It turned out that <i>E. coli</i> has two distantly
related forms of this enzyme, which shows specificity, respectively, for
mononucleotides of nicotinic acid (NadR_2) and nicotinamide (NadD) [<a href="/books/n/sef/A727/?report=reader#A1007">279</a>]. Most other organisms encode either
one or the other form (<a class="figpopup" href="/books/NBK20266/figure/A494/?report=objectonly" target="object" rid-figpopup="figA494" rid-ob="figobA494">Figure 7.14</a>).
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA494" co-legend-rid="figlgndA494"><a href="/books/NBK20266/figure/A494/?report=objectonly" target="object" title="Figure 7.14" class="img_link icnblk_img figpopup" rid-figpopup="figA494" rid-ob="figobA494"><img class="small-thumb" src="/books/NBK20266/bin/ch7f14.gif" src-large="/books/NBK20266/bin/ch7f14.jpg" alt="Figure 7.14. Distribution of NAD biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA494"><h4 id="A494"><a href="/books/NBK20266/figure/A494/?report=objectonly" target="object" rid-ob="figobA494">Figure 7.14</a></h4><p class="float-caption no_bottom_margin">Distribution of NAD biosynthesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A495"><h3>7.5.4. Biotin</h3><p>As is the case with many other pathways, the initial steps of biotin biosynthesis
are poorly understood. The phyletic patterns of the four enzymes that catalyze
the conversion of pimeloyl-CoA into biotin are relatively consistent (<a class="figpopup" href="/books/NBK20266/figure/A496/?report=objectonly" target="object" rid-figpopup="figA496" rid-ob="figobA496">Figure 7.15</a>), but the mechanisms of the
formation of pimelate (6-carboxyhexanoate) and pimeloyl-CoA are still largely
obscure. <i>B. subtilis</i>, <i>A. aeolicus</i>, and
<i>M. jannaschii</i> encode an enzyme that makes pimeloyl-CoA from
pimelate and CoA in a reaction that uses the energy of ATP hydrolysis to AMP and
pyrophosphate [<a href="/books/n/sef/A727/?report=reader#A1403">675</a>]. In contrast,
pimeloyl-CoA synthetase from <i>Pseudomonas mendocina</i> belongs to
the family of NDP-forming acyl-CoA synthetases [<a href="/books/n/sef/A727/?report=reader#A819">91</a>,<a href="/books/n/sef/A727/?report=reader#A1466">738</a>]. Neither of these
two enzyme families is represented in <i>Synechocystis</i> sp.,
<i>H. influenzae</i>, <i>H. pylori</i>, <i>C.
jejuni</i>, and several other bacteria, indicating the existence of yet
another enzyme for the synthesis of pimeloyl-CoA (or an entirely different
pathway for the formation of 7-keto-8-aminopelargonate).
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA496" co-legend-rid="figlgndA496"><a href="/books/NBK20266/figure/A496/?report=objectonly" target="object" title="Figure 7.15" class="img_link icnblk_img figpopup" rid-figpopup="figA496" rid-ob="figobA496"><img class="small-thumb" src="/books/NBK20266/bin/ch7f15.gif" src-large="/books/NBK20266/bin/ch7f15.jpg" alt="Figure 7.15. Distribution of biotin biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA496"><h4 id="A496"><a href="/books/NBK20266/figure/A496/?report=objectonly" target="object" rid-ob="figobA496">Figure 7.15</a></h4><p class="float-caption no_bottom_margin">Distribution of biotin biosynthesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div><p>In spite of the similarity between the phyletic patterns of BioF, BioA, BioD, and
BioB, one cannot help noticing that <i>Synechocystis</i> sp. lacks the
<i>bioA</i> gene, suggesting that amination of
7-keto-8-aminopelargonate is catalyzed by a different aminotransferase. The
absence of <i>bioD</i> and <i>bioB</i> genes in <i>D.
radiodurans</i> makes one wonder whether this bacterium can synthesize
biotin at all.</p><p>The enzyme catalyzing the last reaction in <a class="figpopup" href="/books/NBK20266/figure/A496/?report=objectonly" target="object" rid-figpopup="figA496" rid-ob="figobA496">Figure
7.15</a>, ligation of biotin to the biotin carboxyl carrier protein (or
domain), has a much broader phyletic distribution than any of the biotin
biosynthesis enzymes. This indicates that <i>A. fulgidus</i>,
<i>Halobacterium</i> sp., <i>Pyrococcus</i> spp.,
<i>L. lactis</i>, <i>S. pyogenes</i>, and many other
organisms that do not have a known pathway of biotin synthesis still can utilize
biotin. Thus, they either have a completely different, unknown biotin synthesis
pathway or import biotin from the environment (however, a biotin transport
system so far has not been identified).</p><p>The paucity of data on the enzymes of biotin biosynthesis and a putative biotin
uptake system should encourage active experimentation in this area. There
definitely are novel enzymes and transporters yet to be discovered.</p></div><div id="A497"><h3>7.5.5. Heme</h3><p>From the comparative-genomic point of view, the heme biosynthesis pathway is
characterized by the following trends (<a class="figpopup" href="/books/NBK20266/figure/A498/?report=objectonly" target="object" rid-figpopup="figA498" rid-ob="figobA498">Figure
7.16</a>): (i) with the single exception of uroporphyrinogen III synthase
(HemD), the enzymes from <i>R. prowazekii</i> and yeast (mitochondria)
have identical phyletic patterns; (ii) all archaea, including the aerobes
<i>A. pernix</i> and <i>Halobacterium</i> sp., produce
siroheme but not protoheme; (iii) non-orthologous displacement is observed in
the downstream steps of the pathway, as opposed to the uniformity of all the
upstream steps down to uroporphirinogen III.
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA498" co-legend-rid="figlgndA498"><a href="/books/NBK20266/figure/A498/?report=objectonly" target="object" title="Figure 7.16" class="img_link icnblk_img figpopup" rid-figpopup="figA498" rid-ob="figobA498"><img class="small-thumb" src="/books/NBK20266/bin/ch7f16.gif" src-large="/books/NBK20266/bin/ch7f16.jpg" alt="Figure 7.16. Distribution of heme biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA498"><h4 id="A498"><a href="/books/NBK20266/figure/A498/?report=objectonly" target="object" rid-ob="figobA498">Figure 7.16</a></h4><p class="float-caption no_bottom_margin">Distribution of heme biosynthesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div><div id="A499"><h3>7.5.6. Pyridoxine</h3><p>We conclude our survey of central metabolic pathways with the pyridoxine
biosynthesis pathway, which, despite recent efforts, is still not completely
understood. The scheme below is drawn based on the <i>E. coli</i> data
[<a href="/books/n/sef/A727/?report=reader#A926">198</a>,<a href="/books/n/sef/A727/?report=reader#A1213">485</a>]. In other organisms, the carbon backbone of the
pyridoxine ring is formed of 4-hydroxythreonine (or its phosphate) and
1-deoxy-D-xylulose (or its phosphate) with the nitrogen supplied by either
glutamate (in the PdxAJ-catalyzed reaction), or glutamine (in the
PDX1,PDX2-catalyzed reaction) [<a href="/books/n/sef/A727/?report=reader#A939">211</a>,<a href="/books/n/sef/A727/?report=reader#A1306">578</a>,<a href="/books/n/sef/A727/?report=reader#A1363">635</a>,<a href="/books/n/sef/A727/?report=reader#A1553">825</a>,<a href="/books/n/sef/A727/?report=reader#A1560">832</a>]. Since 1-deoxy-D-xylulose phosphate
synthetase (Dxs, COG1154) so far has been identified only in bacteria, it is
possible that archaea and eukaryotes use a different sugar as a pyridoxine
precursor. Obviously, new enzymes of this pathway remain to be discovered.</p><p>
</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figA500" co-legend-rid="figlgndA500"><a href="/books/NBK20266/figure/A500/?report=objectonly" target="object" title="Figure 7.17" class="img_link icnblk_img figpopup" rid-figpopup="figA500" rid-ob="figobA500"><img class="small-thumb" src="/books/NBK20266/bin/ch7f17.gif" src-large="/books/NBK20266/bin/ch7f17.jpg" alt="Figure 7.17. Distribution of pyridoxine biosynthesis enzymes in organisms with completely sequenced genomes." /></a><div class="icnblk_cntnt" id="figlgndA500"><h4 id="A500"><a href="/books/NBK20266/figure/A500/?report=objectonly" target="object" rid-ob="figobA500">Figure 7.17</a></h4><p class="float-caption no_bottom_margin">Distribution of pyridoxine biosynthesis enzymes in organisms with
completely sequenced genomes. All details are as in Figure
2.7. </p></div></div></div></div><div id="A501"><h2 id="_A501_">7.6. Microbial Enzymes as Potential Drug Targets</h2><p>One of the major incentives behind the genome sequencing of numerous pathogenic
bacteria is the desire to better understand their peculiarities and to develop new
approaches for controlling human diseases caused by these organisms. This task has
become even more urgent with the rapid evolution of antibiotic resistance in many
bacterial pathogens, including multidrug-resistant enterococci, pneumococci,
pseudomonads, staphylococci, and tuberculosis bacilli. Unfortunately, finding new
antibiotics is an extremely laborious process that includes (i) testing numerous
compounds for their activity against model organisms (<i>E. coli</i>,
<i>P. aeruginosa</i>, <i>S. aureus</i>) that are easy to
maintain in culture; (ii) screening these compounds against mammalian cell cultures
to eliminate those toxic to humans; (iii) testing the efficacy and safety of each
chosen drug in animal models; and (iv) pre-clinical and clinical testing, which
alone takes several years. This process ensures that only highly effective and
reasonably safe drugs make it to the market. The majority of drug candidates fail
the tests, usually because in low concentrations they turn out to be safe but
ineffective, whereas at high doses they are effective but show unfavorable side
effects.</p><p>In spite of what one might have read in popular press, genomics cannot accelerate
most steps of the drug development process. What it can do, however, is to increase
the success rate by helping to choose drug candidates that are most likely to be
effective (being targeted at essential systems of the bacterial cell) and least
likely to be toxic (having no targets in the human cell). Indeed, while not all
currently used antibiotics have well-characterized targets, those targets that have
been characterized comprise bacterial proteins that (i) are essential for bacterial
cell metabolism and (ii) are not represented (or represented in a very distinct
form) in human cells (see <a href="/books/n/sef/A298/?report=reader#A319">Table 6.2</a>).
Microbial genome sequences provide us with complete lists of the proteins encoded in
any given pathogen, including all the virulence factors that it could potentially
produce. This &#x0201c;parts list&#x0201d; offers a wide selection of potential
drug targets.</p><p>Comparative analysis of microbial genomes based on the notion of a phyletic pattern,
which is discussed throughout this book, allows the identification of gene products
that are common to all (or most) pathogenic microorganisms in a chosen group, as
well as of those specific for a particular organism. The proteins in the former set
are attractive targets for broad-spectrum antibiotics, whereas the unique proteins
offer an opportunity to design &#x0201c;magic bullets&#x0201d;, which would
specifically target a narrow group of bacteria or even one particular pathogen
[<a href="/books/n/sef/A727/?report=reader#A994">266</a>].</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figA506"><a href="/books/NBK20266/table/A506/?report=objectonly" target="object" title="Table 7.1" class="img_link icnblk_img figpopup" rid-figpopup="figA506" rid-ob="figobA506"><img class="small-thumb" src="/books/NBK20266/table/A506/?report=thumb" src-large="/books/NBK20266/table/A506/?report=previmg" alt="Table 7.1. Cellular targets of most commonly used antibiotics." /></a><div class="icnblk_cntnt"><h4 id="A506"><a href="/books/NBK20266/table/A506/?report=objectonly" target="object" rid-ob="figobA506">Table 7.1</a></h4><p class="float-caption no_bottom_margin">Cellular targets of most commonly used antibiotics. </p></div></div><p>In addition to the lists of probable essential genes, search for potential drug
targets in microbial genomes heavily relies on the understanding of bacterial
metabolism, which is briefly discussed above.</p><div id="A502"><h3>7.6.1. Potential targets for broad-spectrum drugs</h3><p>The list of probable essential genes that potentially could be used as targets
for broad-range antibiotics can be derived using more or less the same approach
as employed for the delineation of the &#x0201c;minimal genome&#x0201d;
([<a href="/books/n/sef/A727/?report=reader#A1180">452</a>], see <a href="/books/n/sef/A22/?report=reader#A40">2.2.5</a>). Inclusion of certain genes in this list is, of
course, affected by non-orthologous gene displacement and enzyme
recruitment.</p><p>It should be noted that compiling the list of the likely essential genes for each
particular group of bacteria (all bacteria, all Gram-positive bacteria, all
mycobacteria, and so on) by computational means is only one of several ways to
accomplish this task, although, arguably, it is the easiest and fastest one. Any
predictions of essentiality for a given gene still have to be verified
experimentally by checking the lethality of knockout mutants [<a href="/books/n/sef/A727/?report=reader#A737">9</a>,<a href="/books/n/sef/A727/?report=reader#A1248">520</a>]. As mentioned above (see <a href="/books/n/sef/A55/?report=reader#A117">3.5.1</a>), lists of essential <i>E. coli</i> genes are
available at <a href="http://www.genome.wisc.edu/resources/essential.htm" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">http://www.genome.wisc.edu/resources/essential.htm</a> and <a href="http://www.shigen.nig.ac.jp/ecoli/pec/Analyses.jsp?key0" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">http://www.shigen.nig.ac.jp/ecoli/pec/Analyses.jsp?key=0</a>.</p><p>In addition to the genes that encode well-characterized essential proteins, the
availability of complete genomes allows one to tap into the pool of
uncharacterized genes whose wide distribution in microbial genomes marks them as
being most likely essential [<a href="/books/n/sef/A727/?report=reader#A785">57</a>,<a href="/books/n/sef/A727/?report=reader#A1179">451</a>]. Searches for such genes can be
easily performed using the &#x0201c;phyletic patterns search&#x0201d; tool
of the COG database. In addition, the COG database contains lists of poorly
characterized and uncharacterized protein families, which are listed as
functional groups R and S, respectively. A collection of uncharacterized
conserved proteins, including those from partially sequenced genomes, is
available in PROSITE database (<a href="http://www.expasy.org/cgi-bin/lists?upflist.txt" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">http://www.expasy.org/cgi-bin/lists?upflist.txt</a>).</p><p>The diversity of microbial metabolic pathways described above offers numerous
possibilities to look for potential drug targets among the metabolic enzymes.
One straightforward approach is to select the pathways that are essential for
certain pathogens but are absent in humans. Such pathways include murein
biosynthesis, the shikimate pathway of aromatic amino acid biosynthesis (<a class="figpopup" href="/books/NBK20266/figure/A452/?report=objectonly" target="object" rid-figpopup="figA452" rid-ob="figobA452">Figure 7.6</a>), and the deoxyxylulose
(non-mevalonate) pathway of terpenoid biosynthesis. It is remarkable that
certain inhibitors of the latter pathway (fosmidomycin, fluoropyruvate,
FR-900098) have been studied as potential antibiotics long before the
characterization of their cellular targets [<a href="/books/n/sef/A727/?report=reader#A1128">400</a>,<a href="/books/n/sef/A727/?report=reader#A1246">518</a>].</p></div><div id="A503"><h3>7.6.2. Potential targets for pathogen-specific drugs</h3><p>Although current approaches favor &#x0201c;one-shot&#x0201d; antibacterials
that can eliminate bacterial infection irrespective of the nature of the
pathogen, it gradually becomes clear that we will soon need a variety of drugs
that would be effective against selected groups of organisms or even a single
pathogen. There is nothing particularly new in this concept: people have been
using anti-tuberculine and anti-syphilis drugs for almost a century without
requiring them to also cure common cold or gastrointestinal problems.</p><p>The novelty stemming from the availability of complete genome sequences is that
now it has become possible to analyze the genome of a pathogen in detail,
looking for weak spots or unusual enzymes that are likely to be essential for
this particular organism. In addition to the traditional drug targets, such as
the cell envelope and the systems for DNA replication, transcription, and
translation, this brings into play for consideration as potential drug targets
such proteins as host interaction factors, transporters for essential nutrients,
enzymes of intermediary metabolism, and many others.</p><p>Host interaction factors can be searched for by using the so-called
&#x0201c;differential genome display&#x0201d;, first proposed by Peer Bork
and his colleagues [<a href="/books/n/sef/A727/?report=reader#A1094">366</a>,<a href="/books/n/sef/A727/?report=reader#A1099">371</a>]. This approach looks for the genes
that are present in the genome of a pathogen but not in the genome of a closely
related free-living bacterium. Because genomes of parasitic bacteria typically
code for fewer proteins than the genomes of their free-living cousins, genes
detected by this approach are likely to be important for pathogenicity. Bork and
colleagues applied this approach to the identification of potential
pathogenicity factors in <i>H. influenzae</i> and <i>H.
pylori</i> through comparison of their genomes against <i>E.
coli</i> [<a href="/books/n/sef/A727/?report=reader#A1094">366</a>,<a href="/books/n/sef/A727/?report=reader#A1099">371</a>].</p><p>Because many pathogens have reduced biosynthetic capabilities and rely on the
host for the supply of certain essential nutrients (see <a href="/books/n/sef/A55/?report=reader#A70">3.2</a>), the respective membrane transport systems can be
valid targets for drug intervention. For example, the still uncharacterized
biotin transport system appears to be the only means of biotin acquisition for
several pathogens, such as <i>S. pyogenes</i>, <i>R.
prowazekii</i>, <i>C. trachomatis</i>, and <i>T.
pallidum</i> (see <a href="#A495">7.5.4</a>).
Actually, one could start probing this hypothetical system right away by using
various biotin analogs. As an added benefit, such a study would eventually lead
to the identification of the transport system components.</p><p>Using surface proteins of bacteria as drug targets has an obvious advantage
because drugs interacting with these proteins do not have to cross the
cytoplasmic membrane, which largely removes the problem of drug efflux-mediated
resistance [<a href="/books/n/sef/A727/?report=reader#A1251">523</a>,<a href="/books/n/sef/A727/?report=reader#A1252">524</a>]. On the other hand, humans also import biotin,
therefore at this stage, it cannot be ruled out that an inhibitor of biotin
uptake might be toxic for humans. This emphasizes the need for identification of
the genes coding for the bacterial uptake system: once these are known, we will
be in a better position to assess the likelihoods of toxic side effects of any
drugs targeting this function.</p><p>Another approach to searching for pathogen-specific drug targets would rely on
the enzymes that are subject to non-orthologous gene displacement and are found
in certain pathogens in a different form that is present in humans. The
rationale for using enzyme inhibitors as antimicrobial drugs comes from the
successful use of sulfamethoxazole and trimethoprim, inhibitors of two different
steps of the folate biosynthetic pathway. Indeed, while each of these drugs is
only moderately effective against most bacterial pathogens, their combination
proved to be effective and reasonably safe. In several instances, detailed
analysis of non-orthologous displacement cases has led to suggestions that
alternative forms of essential enzymes could be used as drug targets ([<a href="/books/n/sef/A727/?report=reader#A994">266</a>,<a href="/books/n/sef/A727/?report=reader#A999">271</a>], see refs. in <a class="figpopup" href="/books/NBK20266/table/A504/?report=objectonly" target="object" rid-figpopup="figA504" rid-ob="figobA504">Table
7.2</a>).</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figA504"><a href="/books/NBK20266/table/A504/?report=objectonly" target="object" title="Table 7.2" class="img_link icnblk_img figpopup" rid-figpopup="figA504" rid-ob="figobA504"><img class="small-thumb" src="/books/NBK20266/table/A504/?report=thumb" src-large="/books/NBK20266/table/A504/?report=previmg" alt="Table 7.2. Examples of pathogen-specific drug targets." /></a><div class="icnblk_cntnt"><h4 id="A504"><a href="/books/NBK20266/table/A504/?report=objectonly" target="object" rid-ob="figobA504">Table 7.2</a></h4><p class="float-caption no_bottom_margin">Examples of pathogen-specific drug targets. </p></div></div></div></div><div id="A505"><h2 id="_A505_">7.7. Conclusions and Outlook</h2><p>This chapter shows that central metabolism is the ultimate playground of
non-orthologous gene displacement, where the logic of phyletic patterns works best.
Metabolic pathways are so amenable to this type of analysis because, if an organism
encodes a significant fraction of the enzymes for a particular pathway, it is
extremely likely that, in reality, is also has the enzymes for the rest of the
steps. Therefore, candidate enzymes for the missing steps may be sought for and, at
least in some instances, found among uncharacterized orthologous sets (identified
through COGs or otherwise) with phyletic patterns that are, at least in part,
complementary to those for known enzymes for the given step. So far, only very few
of the computational predictions made by this approach have been tested
experimentally, but in those studies that have been conducted, the success rate has
been quite high. Conversely, there are enigmatic cases where most of the enzymes of
a given pathway are missing in an organism but one or two still stay around (by
using this language, we imply loss of a pathway, which is indeed largely the case in
parasites and heterotrophs, including ourselves). Most likely, these are cases of
exaptation, where an enzyme that is no longer needed in its original metabolic
capacity has found another job, thus saving itself from extinction. Elucidation of
these exapted functions seems to be an interesting avenue of research.</p><p>The finding that metabolic pathways are so prone to non-orthologous gene displacement
seems to indirectly convey a message of general biological significance. We know for
a fact that enzymes in the same metabolic pathway are connected through reaction
intermediates, but, on almost all occasions, precious little is known about the
actual macromolecular organization of these enzymes in the cell. Analysis of
phyletic patterns shows that many, if not most, metabolic enzymes with different
structures but the same reaction chemistry are interchangeable in evolution. This
suggests that, most of the time, the chemistry is, after all, the principal aspect
of the metabolic functions, whereas the role of co-adaptation of subunits of
macromolecular complexes is likely to be limited.</p><p>The major contribution of lineage-specific gene loss to the evolution of metabolic
pathways is beyond doubt. Horizontal gene transfer is harder to demonstrate but,
realistically, it appears certain that this phenomenon also had a substantial role.
Indeed, it defies credibility to postulate that LUCA had each one of the alternative
forms of metabolic enzymes (and the corresponding reaction intermediates), the
existence of which became apparent through the comparative-genomic studies (as well
as those, perhaps numerous ones that remain to be discovered). The relative
contributions of gene loss and horizontal transfer hopefully will be better
understood through the application of algorithmic methods briefly outlined in <a href="/books/n/sef/A298/?report=reader">Chapter 6</a>.</p><p>Identification of potential targets for antibacterial drugs using phyletic patterns,
the differential genome display technique and other similar approaches is a natural
task for comparative genomics and will likely remain one of its most important
practical applications for years to come.</p></div><div id="A507"><h2 id="_A507_">7.8. Further Reading</h2><dl class="temp-labeled-list"><dl class="bkr_refwrap"><dt>1.</dt><dd><div class="bk_ref" id="A509">Romano AH, Conway T. Evolution of carbohydrate metabolic
pathways. <span><span class="ref-journal">Research in Microbiology. </span>1996;<span class="ref-vol">147</span>:448&ndash;455.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/9084754" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 9084754</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>2.</dt><dd><div class="bk_ref" id="A510">Galperin MY, Walker DR, Koonin EV. Analogous enzymes: independent inventions in enzyme
evolution. <span><span class="ref-journal">Genome Research. </span>1998;<span class="ref-vol">8</span>:779&ndash;790.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/9724324" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 9724324</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>3.</dt><dd><div class="bk_ref" id="A511">Dandekar T, Schuster S, Snel B, Huynen M, Bork P. Pathway alignment: application to the comparative
analysis of glycolytic enzymes. <span><span class="ref-journal">Biochemical Journal. </span>1999;<span class="ref-vol">343</span>:115&ndash;124.</span> [<a href="/pmc/articles/PMC1220531/" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pmc">PMC free article<span class="bk_prnt">: PMC1220531</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/10493919" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10493919</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>4.</dt><dd><div class="bk_ref" id="A512">Huynen MA, Dandekar T, Bork P. Variation and evolution of the citric-acid cycle: a
genomic perspective. <span><span class="ref-journal">Trends in Microbiology. </span>1999;<span class="ref-vol">7</span>:281&ndash;291.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10390638" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10390638</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>5.</dt><dd><div class="bk_ref" id="A513">Cordwell SJ. Microbial genomes and &#x0201c;missing&#x0201d;
enzymes: redefining biochemical pathways. <span><span class="ref-journal">Archives of Microbiology. </span>1999;<span class="ref-vol">172</span>:269&ndash;279.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10550468" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10550468</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>6.</dt><dd><div class="bk_ref" id="A514">Galperin MY, Koonin EV. 2001. Comparative
genome analysis. In: <em>Bioinformatics: a practical guide to the
analysis of genes and proteins</em> (Baxevanis AD and Ouellette
BFF, eds) pp. 359&#x02013;392. John Wiley &#x00026; Sons, New
York. [<a href="https://pubmed.ncbi.nlm.nih.gov/11449732" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 11449732</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>7.</dt><dd><div class="bk_ref" id="A515">Canback B, Andersson SG, Kurland CG. The global phylogeny of glycolytic
enzymes. <span><span class="ref-journal">Proceedings of the National Academy of Sciences of the United
States of America. </span>2002;<span class="ref-vol">99</span>:6097&ndash;6102.</span> [<a href="/pmc/articles/PMC122908/" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pmc">PMC free article<span class="bk_prnt">: PMC122908</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/11983902" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 11983902</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>8.</dt><dd><div class="bk_ref" id="A516">Galperin MY, Koonin EV. Searching for drug targets in microbial
genomes. <span><span class="ref-journal">Current Opinion in Biotechnology. </span>1999;<span class="ref-vol">10</span>:571&ndash;578.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10600691" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10600691</span></a>]</div></dd></dl></dl></div><div style="display:none"><div id="figA520"><img alt="Image ch8f1" src-large="/books/n/sef/A517/bin/ch8f1.jpg" /></div><div id="figA35"><img alt="Image ch2f7" src-large="/books/n/sef/A22/bin/ch2f7.jpg" /></div><div id="figA523"><img alt="Image ch8f2" src-large="/books/n/sef/A517/bin/ch8f2.jpg" /></div></div><div id="bk_toc_contnr"></div></div></div><div class="fm-sec"><h2 id="_NBK20266_pubdet_">Publication Details</h2><h3>Copyright</h3><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright</a> &#x000a9; 2003, Kluwer Academic.</div></div><h3>Publisher</h3><p><a href="http://www.springer.com/" ref="pagearea=page-banner&amp;targetsite=external&amp;targetcat=link&amp;targettype=publisher">Kluwer Academic</a>, Boston</p><h3>NLM Citation</h3><p>Koonin EV, Galperin MY. Sequence - Evolution - Function: Computational Approaches in Comparative Genomics. Boston: Kluwer Academic; 2003. Chapter 7, Evolution of Central Metabolic Pathways: The Playground of Non-Orthologous Gene Displacement.<span class="bk_cite_avail"></span></p></div><div class="small-screen-prev"><a href="/books/n/sef/A298/?report=reader"><svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 100 100" preserveAspectRatio="none"><path d="M75,30 c-80,60 -80,0 0,60 c-30,-60 -30,0 0,-60"></path><text x="20" y="28" textLength="60" style="font-size:25px">Prev</text></svg></a></div><div class="small-screen-next"><a href="/books/n/sef/A517/?report=reader"><svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 100 100" preserveAspectRatio="none"><path d="M25,30c80,60 80,0 0,60 c30,-60 30,0 0,-60"></path><text x="20" y="28" textLength="60" style="font-size:25px">Next</text></svg></a></div></article><article data-type="fig" id="figobA378"><div id="A378" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f1.jpg" alt="Figure 7.1. Distribution of glycolysis (Embden-Meyerhoff-Parnas pathway) enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.1</span><span class="title">Distribution of glycolysis (Embden-Meyerhoff-Parnas pathway)
enzymes in organisms with completely sequenced genomes</span></h3><div class="caption"><p> Each rounded rectangle shows a glycolytic enzyme, denoted by its
gene name and the COG number. Alternative enzymes catalyzing the
same reaction are shown side-by-side. Each COG is accompanied by
its phyletic pattern (see <a href="/books/n/sef/A22/?report=reader#A43">2.2.6</a>). The species abbreviations are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure 2.7</a>.</p></div></div></article><article data-type="fig" id="figobA386"><div id="A386" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f2.jpg" alt="Figure 7.2. Distribution of gluconeogenesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.2</span><span class="title">Distribution of gluconeogenesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA391"><div id="A391" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f3.jpg" alt="Figure 7.3. Distribution of enzymes of the pentose phosphate and Entner-Doudoroff pathways in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.3</span><span class="title">Distribution of enzymes of the pentose phosphate and
Entner-Doudoroff pathways in organisms with completely sequenced
genomes</span></h3><div class="caption"><p>Details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA411"><div id="A411" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f4.jpg" alt="Figure 7.4. Distribution of the TCA cycle enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.4</span><span class="title">Distribution of the TCA cycle enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA431"><div id="A431" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f5.jpg" alt="Figure 7.5. Distribution of purine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.5</span><span class="title">Distribution of purine biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details as in <a href="/books/n/sef/A22/?report=reader#A35">Figure 2.7</a>.</p></div></div></article><article data-type="fig" id="figobA452"><div id="A452" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f6.jpg" alt="Figure 7.6. Distribution of tryptophan biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.6</span><span class="title">Distribution of tryptophan biosynthesis enzymes in
organisms with completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA468"><div id="A468" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f7.jpg" alt="Figure 7.7. Distribution of phenylalanine and tyrosine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.7</span><span class="title">Distribution of phenylalanine and tyrosine biosynthesis
enzymes in organisms with completely sequenced
genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA474"><div id="A474" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f8.jpg" alt="Figure 7.8. Distribution of arginine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.8</span><span class="title">Distribution of arginine biosynthesis enzymes in organisms
with completely sequenced genomes</span></h3><div class="caption"><p>All details as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA483"><div id="A483" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f9.jpg" alt="Figure 7.9. Distribution of histidine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.9</span><span class="title">Distribution of histidine biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA485"><div id="A485" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f10.jpg" alt="Figure 7.10. Distribution of isoleucine/leucine/valine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.10</span><span class="title">Distribution of isoleucine/leucine/valine biosynthesis enzymes in
organisms with completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA487"><div id="A487" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f11.jpg" alt="Figure 7.11. Distribution of proline biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.11</span><span class="title">Distribution of proline biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3></div></article><article data-type="fig" id="figobA490"><div id="A490" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f12.jpg" alt="Figure 7.12. Distribution of thiamine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.12</span><span class="title">Distribution of thiamine biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA492"><div id="A492" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f13.jpg" alt="Figure 7.13. Distribution of riboflavin biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.13</span><span class="title">Distribution of riboflavin biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA494"><div id="A494" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f14.jpg" alt="Figure 7.14. Distribution of NAD biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.14</span><span class="title">Distribution of NAD biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA496"><div id="A496" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f15.jpg" alt="Figure 7.15. Distribution of biotin biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.15</span><span class="title">Distribution of biotin biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA498"><div id="A498" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f16.jpg" alt="Figure 7.16. Distribution of heme biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.16</span><span class="title">Distribution of heme biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="fig" id="figobA500"><div id="A500" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK20266/bin/ch7f17.jpg" alt="Figure 7.17. Distribution of pyridoxine biosynthesis enzymes in organisms with completely sequenced genomes." /></div><h3><span class="label">Figure 7.17</span><span class="title">Distribution of pyridoxine biosynthesis enzymes in organisms with
completely sequenced genomes</span></h3><div class="caption"><p>All details are as in <a href="/books/n/sef/A22/?report=reader#A35">Figure
2.7</a>.</p></div></div></article><article data-type="table-wrap" id="figobA506"><div id="A506" class="table"><h3><span class="label">Table 7.1</span><span class="title">Cellular targets of most commonly used antibiotics</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK20266/table/A506/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__A506_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">
Antibiotic groups, Examples
</th><th id="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">
Bacterial target
</th><th id="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">
Resistance mechanisms
</th></tr></thead><tbody><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">&#x003b2;<b>-Lactams:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Penicillins, cephalosporins,
carbapenems, monobactams</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Peptidoglycan transpeptidase, other
proteins</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Hydrolysis by &#x003b2;-lactamase,
alterations in penicillin-binding proteins</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Glycopeptides:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Bacitracin, colistin, dactinomycin,
teichoplanin, vancomycin, virginiamycin</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Peptidoglycan transpeptidase,
transglycosylase</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Modification of the UDP-muramyl
pentapeptide</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Aminoglycosides:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Amikacin, kanamycin, gentamycin,
hygromycin, neomycin, puromycin, streptomycin, tobramycin</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Ribosomal 30S subunit</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Acetylation, adenylation, or
phosphorylation of the antibiotic by specific modifying
enzymes</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Tetracyclins:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Doxycycline, methacycline,
minocycline, tetracycline</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Ribosomal 30S subunit</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Export by efflux pumps, mutations</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Macrolides:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Azithromycin, dirithromycin,
clarithromycin, spiramycin, erythromycin, oleandomycin</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Ribosomal 50S subunit</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">rRNA methylation;
<i>rrn</i>, <i>rplD</i>, and
<i>rplV</i> mutations; hydrolysis by esterases,
export by efflux pumps</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Quinolones:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Nalidixic acid, ciprofloxacin</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">DNA gyrase &#x003b2;-subunit</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>gyrB</i> mutations</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Lincosamides:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Clindamycin, lincomycin</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">23S rRNA</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">rRNA methylation, <i>rrn</i>
mutations, drug adenylation</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Chloramphenicol</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Peptidyl-transferase center on the
ribosomal 50S subunit</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Inactivation by acetylation, export by
efflux pumps</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Sulfonamides:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Sulfamethoxazole</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Dihydropteroate synthase</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>folP</i> mutations</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Trimethoprim</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Dihydrofolate reductase</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>folA</i> mutations</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Nitroimidazoles:</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Metronidazole</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Chromosomal DNA</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Nitroreductase mutations, preventing
drug activation</td></tr><tr><td headers="hd_h_A506_1_1_1_1 hd_h_A506_1_1_1_2 hd_h_A506_1_1_1_3" colspan="3" rowspan="1" style="vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<b>Rifampin</b>
</td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td headers="hd_h_A506_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_A506_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">RNA polymerase &#x003b2;-subunit</td><td headers="hd_h_A506_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>rpoB</i> mutations</td></tr></tbody></table></div></div></article><article data-type="table-wrap" id="figobA504"><div id="A504" class="table"><h3><span class="label">Table 7.2</span><span class="title">Examples of pathogen-specific drug targets</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK20266/table/A504/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__A504_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
Enzymes with limited phyletic distribution
</th><th id="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
Human pathogens that depend on these enzymes
</th><th id="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
Ref.
</th></tr></thead><tbody><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">ATP/ADP translocase, bacterial/plant
type</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>R. prowazekii, C. trachomatis, C. pneumoniae</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A1623">895</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">3-Dehydroquinate dehydratase, class
II</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>C. jejuni</i>, <i>H. influenzae</i>,
<i>H. pylori</i>, <i>P. aeruginosa</i>,
<i>V. cholerae</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A1033">305</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">DhnA-type fructose-1,6-bisphosphate
aldolase</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>C. trachomatis</i>, <i>C. pneumoniae</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A985">257</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Lysyl-tRNA synthetase, class I</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>B. burgdorferi, R. prowazekii, T. pallidum,</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A1103">375</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Na-translocating NADH: ubiquinone
oxidoreductase</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>C. trachomatis, C. pneumoniae, Cl. perfringens, T.
denticola</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A1062">334</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Na-translocating oxalo-acetate
decarboxylase</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>S. pyogenes, T. pallidum,</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A1062">334</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Orotidine 5&#x02032;-phosphate
decarboxylase</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>M. leprae, M. tuberculosis</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A740">12</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Pyridoxine biosynthesis enzymes PDX1,
PDX2</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>Bacillus anthracis, H. influenzae, L. monocytogenes, M.
leprae, M. tuberculosis, S. pneumoniae</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A991">263</a>,<a href="/books/n/sef/A727/?report=reader#A1363">635</a>]</td></tr><tr><td headers="hd_h_A504_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Cofactor-independent phosphoglycerate
mutase</td><td headers="hd_h_A504_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
<i>C. jejuni, H. pylori, M. genitalium, P.
aeruginosa</i>, <i>V. cholerae</i>
</td><td headers="hd_h_A504_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">[<a href="/books/n/sef/A727/?report=reader#A986">258</a>,<a href="/books/n/sef/A727/?report=reader#A989">261</a>]</td></tr></tbody></table></div></div></article></div><div id="jr-scripts"><script src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/libs.min.js"> </script><script src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/jr.min.js"> </script></div></div>
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