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<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="citation_inbook_title" content="Probe Reports from the NIH Molecular Libraries Program [Internet]" /><meta name="citation_title" content="Probe Report for PME-1 Inhibitors" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2010/10/20" /><meta name="citation_author" content="DA Bachovchin" /><meta name="citation_author" content="AE Speers" /><meta name="citation_author" content="AM Zuhl" /><meta name="citation_author" content="SJ Brown" /><meta name="citation_author" content="BF Cravatt" /><meta name="citation_author" content="Fernandez-Vega, V" /><meta name="citation_author" content="T Spicer" /><meta name="citation_author" content="BA Mercer" /><meta name="citation_author" content="J Ferguson" /><meta name="citation_author" content="P Hodder" /><meta name="citation_author" content="HR Rosen" /><meta name="citation_pmid" content="21433379" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK50686/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Probe Report for PME-1 Inhibitors" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="DA Bachovchin" /><meta name="DC.Contributor" content="AE Speers" /><meta name="DC.Contributor" content="AM Zuhl" /><meta name="DC.Contributor" content="SJ Brown" /><meta name="DC.Contributor" content="BF Cravatt" /><meta name="DC.Contributor" content="Fernandez-Vega, V" /><meta name="DC.Contributor" content="T Spicer" /><meta name="DC.Contributor" content="BA Mercer" /><meta name="DC.Contributor" content="J Ferguson" /><meta name="DC.Contributor" content="P Hodder" /><meta name="DC.Contributor" content="HR Rosen" /><meta name="DC.Date" content="2010/10/20" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK50686/" /><meta name="description" content="Recent findings have identified protein phosphatase methylesterase-1 (PME-1) as a protector of sustained ERK pathway activity in malignant gliomas. PME-1 is a protein methylesterase that functions in the regulation of protein phosphatase 2A (PP2A) by reversible methylation. Biochemical elucidation of PME-1 would thus greatly benefit from the development of potent and selective chemical inhibitors. The probe compound ML136 (CID-44607965), containing a sulfonyl acrylonitrile core, represents the first potent, selective inhibitor of PME-1. Moreover, the probe does not appear to exhibit cytotoxicity. Thus, ML136 should serve as a useful tool for in vitro and in situ research assays in which it is desirable to specifically block PME-1 activity." /><meta name="og:title" content="Probe Report for PME-1 Inhibitors" /><meta name="og:type" content="book" /><meta name="og:description" content="Recent findings have identified protein phosphatase methylesterase-1 (PME-1) as a protector of sustained ERK pathway activity in malignant gliomas. PME-1 is a protein methylesterase that functions in the regulation of protein phosphatase 2A (PP2A) by reversible methylation. Biochemical elucidation of PME-1 would thus greatly benefit from the development of potent and selective chemical inhibitors. The probe compound ML136 (CID-44607965), containing a sulfonyl acrylonitrile core, represents the first potent, selective inhibitor of PME-1. Moreover, the probe does not appear to exhibit cytotoxicity. 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<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>Probe Reports from the NIH Molecular Libraries Program [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2010-. </p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/mlprobe/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-mlprobe-lrg.png" alt="Cover of Probe Reports from the NIH Molecular Libraries Program" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>Probe Reports from the NIH Molecular Libraries Program [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK50686_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK50686_dtls__"><div>Bethesda (MD): National Center for Biotechnology Information (US); 2010-.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/mlprobe/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/mlprobe/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/mlprobe/ml137/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/mlprobe/ml135/" title="Next page in this title">Next ></a></div></div></div></div></div>
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<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK50686_"><span class="title" itemprop="name">Probe Report for PME-1 Inhibitors</span></h1><p class="contrib-group"><span itemprop="author">DA Bachovchin</span>, <span itemprop="author">AE Speers</span>, <span itemprop="author">AM Zuhl</span>, <span itemprop="author">SJ Brown</span>, <span itemprop="author">BF Cravatt</span>, <span itemprop="author">Fernandez-Vega, V</span>, <span itemprop="author">T Spicer</span>, <span itemprop="author">BA Mercer</span>, <span itemprop="author">J Ferguson</span>, <span itemprop="author">P Hodder</span>, and <span itemprop="author">HR Rosen</span>.</p><p class="small">Received: <span itemprop="datePublished">February 26, 2010</span>; Last Update: <span itemprop="dateModified">October 20, 2010</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="_abs_rndgid_" itemprop="description"><p>Recent findings have identified protein phosphatase methylesterase-1 (PME-1) as a protector of sustained ERK pathway activity in malignant gliomas. PME-1 is a protein methylesterase that functions in the regulation of protein phosphatase 2A (PP2A) by reversible methylation. Biochemical elucidation of PME-1 would thus greatly benefit from the development of potent and selective chemical inhibitors. The probe compound ML136 (CID-44607965), containing a sulfonyl acrylonitrile core, represents the first potent, selective inhibitor of PME-1. Moreover, the probe does not appear to exhibit cytotoxicity. Thus, ML136 should serve as a useful tool for in vitro and in situ research assays in which it is desirable to specifically block PME-1 activity.</p></div><div class="h2"></div><p><b>Assigned Assay Grant #:</b> CA132630-01</p><p><b>Screening Center Name & PI:</b> Scripps Research Institute Molecular Screening Center (SRIMSC), H. Rosen</p><p><b>Chemistry Center Name & PI:</b> SRIMSC, H. Rosen</p><p><b>Assay Submitter & Institution:</b> Ben Cravatt, TSRI</p><p><b>PubChem Summary Bioassay Identifier (AID):</b>
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<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2143" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2143</a></p><div id="ml136.fu1" class="figure"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu1.jpg" alt="Image ml136fu1" /></div></div><div id="ml136.tu1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu1_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b>PME-1 Inhibitor Probe</b></td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b>(Sulfonyl Acrylonitrile Scaffold)</b></td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID 44607965 <a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457340" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-87457340</a></td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">MLS002699139</td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b>PME-1 IC</b><sub><b>50</b></sub>: 0.5 μM</td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b>Anti-Target (other serine hydrolases) IC</b><sub><b>50</b></sub>:>100 μM</td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b>Cytox CC</b><sub><b>50</b></sub>: >100 μM</td></tr><tr><td rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b><a href="/pcsubstance/?term=ML136[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML136</a></b></td></tr></tbody></table></div></div><div id="ml136.s2"><h2 id="_ml136_s2_">Probe Structure & Characteristics</h2><p>As described in the CPDP, the chief goal for this probe development project was to find a selective inhibitor of the protein methylesterase PME-1. The probe compound <a href="/pcsubstance/?term=ML136[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML136</a> (CID 44607965), having a sulfonyl acrylonitrile core, is claimed as a potent and selective inhibitor of PME-1. This probe is highly (>40-fold) selective among other serine hydrolases, as demonstrated by activity in the gel-based proteomic profiling assay. In addition, the probe compound CID 44607965 exhibits no cytotoxicity. This compound is the first selective inhibitor of PME-1.</p><p>Please see the probe table on the next page for all results described above.</p></div><div id="ml136.s3"><h2 id="_ml136_s3_">Probe Selection</h2><p>Following the uHTS campaign and counterscreening by gel-based competitive activity-based protein profiling (ABPP) in proteomes, a sulfonyl acrylonitrile class of inhibitor was identified for probe development. Following two rounds of SAR, compound CID 44607965 was selected as a probe since it was the most potent and selective compound tested for in situ studies. It was ~20-fold more potent than the top initial hit (CID 5703330) and at least 20-fold more selective.</p></div><div id="ml136.s4"><h2 id="_ml136_s4_">Recommendations for the Scientific Use of This Probe</h2><p>This compound is useful for in vitro or in situ research assays in which it is desirable to specifically block PME-1 activity.</p></div><div id="ml136.s5"><h2 id="_ml136_s5_">1. Scientific Rationale for Project</h2><p>Reversible protein phosphorylation networks play essential roles in most cellular processes. While over 500 kinases catalyze protein phosphorylation, only two enzymes, PP1 and PP2A, are responsible for >90% of all serine/threonine phosphatase activity[<a class="bk_pop" href="#ml136.r1">1</a>]. Phosphatases, unlike kinases, achieve substrate specificity through complex subunit assembly and post-translational modifications rather than number. PP2A, for example, typically exists as heterotrimer with diverse subunits that may combinatorially make as many as 70 different holoenzyme assemblies[<a class="bk_pop" href="#ml136.r2">2</a>]. Mutations in several of these PP2A subunits have been identified in human cancers, suggesting that PP2A may act as a tumor suppressor[<a class="bk_pop" href="#ml136.r3">3</a>]. Adding further complexity, several residues of the catalytic subunit of PP2A can be reversibly phosphorylated, and the C-terminal leucine residue can be reversibly methylated [<a class="bk_pop" href="#ml136.r4">4</a>, <a class="bk_pop" href="#ml136.r5">5</a>]. PME-1 is specifically responsible for demethylation of the carboxyl terminus [<a class="bk_pop" href="#ml136.r4">4</a>].</p><p>Methylesterification is thought to control the binding of different subunits to PP2A, but little is known about physiological significance of this post-translational modification <i>in vivo</i> [<a class="bk_pop" href="#ml136.r5">5</a>]. Recently, PME-1 has been identified as a protector of sustained ERK pathway activity in malignant gliomas [<a class="bk_pop" href="#ml136.r6">6</a>]. In order to further elucidate the role of PP2A methylation <i>in vivo</i>, our lab has generated mice that lack PME-1 (PME-1 knockout mice) by targeted gene disruption [<a class="bk_pop" href="#ml136.r7">7</a>]. Unfortunately, <i>PME-1</i> deletion resulted in perinatal lethality, underscoring the importance of PME-1 but hindering our biological studies. Biochemical elucidation of PME-1 would thus greatly benefit from the development of potent and selective chemical inhibitors.</p><p>As a serine hydrolase, catalytically active PME-1 is readily labeled by fluorescent activity-based protein profiling (ABPP) probes bearing a fluorophosphonate (FP) reactive group [<a class="bk_pop" href="#ml136.r8">8</a>]. This reactivity can be exploited for inhibitor discovery using a competitive-ABPP platform, whereby small molecule enzyme inhibition is assessed by the ability to out-compete ABPP probe labeling [<a class="bk_pop" href="#ml136.r9">9</a>]. When used in the context of a complex proteome, competitive-ABPP also offers a means to assess inhibitor selectivity against a wide range of probe-reactive enzymes. Competitive ABPP has also been configured to operate in a high-throughput manner via fluorescence polarization readout, FluoPol-ABPP [<a class="bk_pop" href="#ml136.r10">10</a>], offering a facile means to screen for PME-1 inhibitors.</p><div id="ml136.tu2" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu2/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu2_lrgtbl__"><table class="no_bottom_margin"><thead><tr><th id="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Probe or Analog</th><th id="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">CID/ML</th><th id="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SID</th><th id="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS</th><th id="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Target Name</th><th id="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Target IC<sub>50</sub> [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2366</a>] (μM)</th><th id="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Anti-target Name</th><th id="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Anti-Target IC<sub>50</sub> [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2366</a>] (μM)</th><th id="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Selectivity</th><th id="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay Cytox CC<sub>50</sub> [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2365" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2365</a>] (nM)</th></tr></thead><tbody><tr><td headers="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Probe</b></td><td headers="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607965/<a href="/pcsubstance/?term=ML136[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML136</a></td><td headers="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457340" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457340</a></td><td headers="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699139</td><td headers="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">0.5</td><td headers="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>30 serine hydrolases (SHs)</td><td headers="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>20</td><td headers="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">>40-fold</td><td headers="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/100000" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">100000</a></td></tr><tr><td headers="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 1</b></td><td headers="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607949</td><td headers="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457341" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457341</a></td><td headers="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699140</td><td headers="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">0.6</td><td headers="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>30 SHs</td><td headers="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>20</td><td headers="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">>33-fold</td><td headers="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/100000" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">100000</a></td></tr><tr><td headers="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 2</b></td><td headers="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607954</td><td headers="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457343" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457343</a></td><td headers="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699141</td><td headers="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">3.0</td><td headers="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>30 SHs</td><td headers="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>20</td><td headers="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">>6-fold</td><td headers="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NT<sup><a class="bk_pop" href="#ml136.tfn1">*</a></sup></td></tr><tr><td headers="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 3</b></td><td headers="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607974</td><td headers="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457347" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457347</a></td><td headers="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699142</td><td headers="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">4.8</td><td headers="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>30 SHs</td><td headers="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>20</td><td headers="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">>4.2-fold</td><td headers="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NT</td></tr><tr><td headers="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 4</b></td><td headers="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607969</td><td headers="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457348" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457348</a></td><td headers="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699143</td><td headers="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">6.8</td><td headers="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>30 SHs</td><td headers="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>20</td><td headers="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">>2-fold</td><td headers="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NT</td></tr><tr><td headers="hd_h_ml136.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 5</b></td><td headers="hd_h_ml136.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5703330</td><td headers="hd_h_ml136.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457336" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457336</a></td><td headers="hd_h_ml136.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699144</td><td headers="hd_h_ml136.tu2_1_1_1_5" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu2_1_1_1_6" rowspan="1" colspan="1" style="background-color:#CCFFFF;text-align:center;vertical-align:middle;">10.8</td><td headers="hd_h_ml136.tu2_1_1_1_7" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">>30 SHs</td><td headers="hd_h_ml136.tu2_1_1_1_8" rowspan="1" colspan="1" style="background-color:#CCFFCC;text-align:center;vertical-align:middle;">20 75kDa SH</td><td headers="hd_h_ml136.tu2_1_1_1_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><2-fold</td><td headers="hd_h_ml136.tu2_1_1_1_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NT</td></tr></tbody></table></div><div><div><dl class="temp-labeled-list small"><dt>*</dt><dd><div id="ml136.tfn1"><p class="no_margin">NT: not tested</p></div></dd></dl></div></div></div></div><div id="ml136.s6"><h2 id="_ml136_s6_">2. Project Description</h2><div id="ml136.s7"><h3>a. Original goal for probe characteristics</h3><p>The goal of the campaign was to discover compounds with inhibitory activity against PME-1 that are selective among the serine hydrolases in mouse tissue and human cell line proteomes as assessed by gel-based ABPP. Compounds of interest should exhibit an IC<sub>50</sub> of <10 μM potency, and preferably <1 μM potency.</p></div><div id="ml136.s8"><h3>b. Information for each Assay Implemented and Screening Run</h3><div id="ml136.s9"><h4>i. PubChem Bioassay Name(s), AID(s), Assay-Type (Primary, DR, Counterscreen, Secondary)</h4><div id="ml136.tu3" class="table"><h3><span class="title">PubChem BioAssay Table</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu3/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu3_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">AID</th><th id="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Assay Name</th><th id="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Assay Type</th><th id="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Target</th><th id="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Powder Sample</th><th id="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Compound Concentration</th></tr></thead><tbody><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2130" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2130</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Primary biochemical high-throughput screening assay to measure PME-1 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Primary Assay (1X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.9 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2268" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2268</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Primary Marybridge biochemical high-throughput screening assay to measure PME-1 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Primary Assay (1X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">8 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2171" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2171</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Confirmation biochemical high-throughput screening assay to measure PME-1 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Confirmation Assay (3X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.9 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2174" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2174</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: Primary biochemical high- throughput screening assay to measure LYPLA1 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Primary Assay (1X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">LYPLA1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.9 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2233" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2233</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: Confirmation biochemical high-throughput screening assay to measure LYPLA1 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Confirmation Assay (3X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">LYPLA1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.9 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2177" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2177</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: Primary biochemical high- throughput screening assay to measure LYPLA2 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Primary Assay (1X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">LYPLA2</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.9 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2232" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2232</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: Confirmation biochemical high-throughput screening assay to measure LYPLA2 inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Confirmation Assay (3X, %INH)</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">LYPLA2</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.9 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2369" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2369</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Late stage results from the probe development effort to identify inhibitors of Protein Methylesterase-1 (PME-1): Gel-based Activity-Based Protein Profiling (ABPP) Inhibition</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Yes</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">20 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2366</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Late stage results from the probe development effort to identify inhibitors of Protein Methylesterase-1 (PME-1): Gel-based Activity-Based Protein Profiling (ABPP) IC<sub>50</sub></td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1, anti-target serine hydrolases</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Yes</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.1 – 100 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2371" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2371</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Late stage results from the probe development effort to identify inhibitors of Protein Methylesterase-1 (PME-1): Gel-based Activity-Based Protein Profiling (ABPP) IC<sub>50</sub>: Purified enzyme</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Yes</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.1 – 100 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2365" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2365</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Late stage results from the probe development effort to identify inhibitors of Protein Methylesterase-1 (PME-1): Luminescence- based counterscreen assay to identify cytotoxic compounds</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">HEK 293T cells</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Yes</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.001 – 100 μM</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2368" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2368</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Late stage results from the probe development effort to identify inhibitors of Protein Methylesterase-1 (PME-1): Gel-based Activity-Based Protein Profiling (ABPP) Gel Filtration Assay</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Yes</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">100 M</td></tr><tr><td headers="hd_h_ml136.tu3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2363" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2363</a></td><td headers="hd_h_ml136.tu3_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Late stage results from the probe development effort to identify inhibitors of Protein Methylesterase-1 (PME-1): Inhibition of PP2A demethylation in HeLa cells</td><td headers="hd_h_ml136.tu3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Secondary Assay</td><td headers="hd_h_ml136.tu3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1</td><td headers="hd_h_ml136.tu3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Yes</td><td headers="hd_h_ml136.tu3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5 μM and 20 μM</td></tr></tbody></table></div></div></div><div id="ml136.s10"><h4>ii. Assay Rationale & Description</h4><div id="ml136.tu4" class="table"><h3><span class="title">Table of Assay Rationale and Description</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu4/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu4_lrgtbl__"><table class="no_margin"><thead><tr><th id="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">AID</th><th id="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Assay Rationale</th><th id="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Assay Description</th><th id="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Z′</th><th id="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">S:B</th></tr></thead><tbody><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2130" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2130</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To measure the ability of compounds to inhibit PME-1 activity</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">In this assay, a fluorophosphonate-rhodamine (FP-Rh) probe which broadly targets enzymes from the serine hydrolase family is used to label PME-1 in the presence of test compounds. The reaction is excited with linear polarized light and the intensity of the emitted light is measured as the polarization value (mP). As designed, test compounds that act as PME-1 inhibitors will prevent PME-1-probe interactions, thereby increasing the proportion of free (unbound) fluorescent probe in the well, leading to low fluorescence polarization in the well. Compounds were tested in singlicate at a final nominal concentration of 5.9 micromolar.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.76</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1.72</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2268" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2268</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To measure the ability of compounds to inhibit PME-1 activity (Maybridge Library)</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Same as above except compounds were tested at 8.0 micromolar.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.78</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">3.17</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2171" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2171</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Confirmation of hit activity of compounds identified in the Primary Screen</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Same as 2130 except compounds were tested in triplicate.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.73</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2174" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2174</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: To measure the ability of compounds to inhibit LYPLA1</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">In this assay, a fluorophosphonate-rhodamine (FP-Rh) probe which broadly targets enzymes from the serine hydrolase family is used to label LYPLA1 in the presence of test compounds. The reaction is excited with linear polarized light and the intensity of the emitted light is measured as the polarization value (mP). As designed, test compounds that act as LYPLA1 inhibitors will prevent LYPLA1-probe interactions, thereby increasing the proportion of free (unbound) fluorescent probe in the well, leading to low fluorescence polarization in the well. Compounds were tested in singlicate at a final nominal concentration of 5.9 micromolar.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.86</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2.12</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2233" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2233</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: Confirmation of activity of compounds identified in 2174</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Same as above except compounds were tested in triplicate.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.89</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2.3</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2177" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2177</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: To measure the ability of compounds to inhibit LYPLA2</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">In this assay, a fluorophosphonate-rhodamine (FP-Rh) probe which broadly targets enzymes from the serine hydrolase family is used to label LYPLA2 in the presence of test compounds. The reaction is excited with linear polarized light and the intensity of the emitted light is measured as the polarization value (mP). As designed, test compounds that act as LYPLA2 inhibitors will prevent LYPLA2-probe interactions, thereby increasing the proportion of free (unbound) fluorescent probe in the well, leading to low fluorescence polarization in the well. Compounds were tested in singlicate at a final nominal concentration of 5.9 micromolar.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.77</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1.69</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2232" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2232</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Counterscreen for PME1 inhibitors: Confirmation of activity of compounds identified in 2177</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Same as above except compounds were tested in triplicate.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.78</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1.69</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2369" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2369</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To confirmation activity of compounds in proteomes</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">In this assay, a fluorophosphonate-rhodamine (FP-Rh) probe which broadly targets enzymes from the serine hydrolase family is used to label PME-1 in mouse brain soluble lysates in the presence of test compounds. The reaction products are separated by SDS-PAGE and visualized in-gel using a flatbed fluorescence scanner. The percentage activity remaining is determined by measuring the integrated optical density of the bands. As designed, test compounds that act as PME-1 inhibitors will prevent PME-1-probe interactions, thereby increasing the proportion of free (unbound) fluorescent probe, leading to low fluorescence intensity in the band in the gel. Percent inhibition of PME- 1 in mouse brain soluble lysates (compound at 20 μM) was determined.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2366</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To determine selectivity of compounds in proteomes</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Same as above except IC<sub>50</sub> values are determined from dose-response curves from three trials at each inhibitor concentration (0.1–100 mM).</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2371" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2371</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To confirm activity ofcompounds with purified enzyme</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Same as above except purified PME-1 (not in lysates) was tested. IC<sub>50</sub> values are determined from dose-response curves from three trials at each inhibitor concentration (0.1–100 mM).</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2365" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2365</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To determine cytotoxicity of inhibitor compounds</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">In this assay, HEK cells are incubated with test compounds, followed by determination of cell viability. The assay utilizes the CellTiter- Glo luminescent reagent to measure intracellular ATP in viable cells. Luciferase present in the reagent catalyzes the oxidation of beetle luciferin to oxyluciferin and light in the presence of cellular ATP. Well luminescence is directly proportional to ATP levels and cell viability. As designed, compounds that reduce cell viability will reduce ATP levels, luciferin oxidation and light production, resulting in decreased well luminescence. Compounds were tested in triplicate in a 7-point 1:10 dilution series starting at a nominal test concentration of 100 micromolar.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2368" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2368</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To assess reversibility of binding of inhibitor compounds</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">In this assay, a fraction of the enzyme-inhibitor mixture is passaged over a Sephadex G-25M column (GE Healthcare) before reaction with a fluorophosphonate-rhodamine (FP-Rh) probe which broadly targets enzymes from the serine hydrolase family. The reaction products are separated by SDS-PAGE and visualized in-gel using a flatbed fluorescence scanner. The percentage activity remaining is determined by measuring the integrated optical density of the bands. As designed, test compounds that act as irreversible PME-1 inhibitors will prevent PME-1-probe interactions after gel filtration, thereby increasing the proportion of free (unbound) fluorescent probe, leading to low fluorescence intensity in the band in the gel. The compound’s reversibility of inhibition of PME-1 was assessed.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td></tr><tr><td headers="hd_h_ml136.tu4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2363" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2363</a></td><td headers="hd_h_ml136.tu4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">To confirm that compounds inhibit the ability of PME-1 to demethylate PP2A in HeLA cells</td><td headers="hd_h_ml136.tu4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">The purpose of this assay is to monitor the demethylation of PP2A by endogenous PME-1 that occurs during inhibitor incubation with cells in culture (e.g., HeLa) at 37 °C. Following incubation with compound, cells are homogenized, and proteins are separated by SDS- PAGE. PP2A demethylation is visualized by chemi-luminescent dectection using HRP-antibody specific to C-terminal demethylated PP2A. As designed, test compounds that act as PME-1 inhibitors will inhibit demethylation of PP2A, resulting in a decrease in the demethylated PP2a signal.</td><td headers="hd_h_ml136.tu4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td><td headers="hd_h_ml136.tu4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N/A</td></tr></tbody></table></div><div><div><dl class="temp-labeled-list small"><dt></dt><dd><div id="ml136.tfn2"><p class="no_margin">N/A: Not applicable.</p></div></dd></dl></div></div></div><div id="ml136.tu5" class="table"><h3><span class="title">Table of Reagents and Source</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu5/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu5_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">AID</th><th id="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Reagent (Source)</th></tr></thead><tbody><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1 Inhibition Assays (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2130" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2130</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2269" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2269</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2171" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2171</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Recombinant PME-1 enzyme (supplied by Assay Provider)<br />FP-Rh probe (supplied by Assay Provider)<br />Tris HCl (Sigma, part T3038)<br />NaCl (Sigma, part S6546)<br />Pluronic acid (Invitrogen, part P6866)<br />1536-well plates (Greiner, part 789176)<br />DTT (Invitrogen 15508-013)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1 Counterscreen Assays: LYPLA1 (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2174" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2174</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2233" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2233</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Recombinant LYPLA1 enzyme (supplied by Assay Provider)<br />Substrate (FP-Rh probe) (supplied by Assay Provider)<br />Tris HCl (Sigma, part T3038)<br />NaCl (Sigma, part S6546)<br />Pluronic acid (Invitrogen, part P6866)<br />1536-well plates (Greiner, part 789176)<br />DTT (Invitrogen, part 15508-013)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1 Counterscreen Assays: LYPLA2 (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2177" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2177</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2232" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2232</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Recombinant LYPLA2 enzyme (supplied by Assay Provider)<br />Substrate (FP-Rh probe) (supplied by Assay Provider)<br />Tris HCl (Sigma, part T3038)<br />NaCl (Sigma, part S6546)<br />Pluronic acid (Invitrogen, part P6866)<br />1536-well plates (Greiner, part 789176)<br />DTT (Invitrogen, part 15508-013)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Cytotoxicity Assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2365" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2365</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">HEK 293T cells<br />Cell Titer-Glo (Promega, part <a href="/nuccore/22474626" class="bk_tag" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=nuccore">G75729</a>)<br />96-well plates</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">ABPP % INH Assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2369" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2369</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Mouse brain cytosol<br />FP-Rh probe<br />Sodium Chloride (Fisher, part 980597)<br />1M Tris, pH 8.0 (Invitrogen, part T-3038)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">ABPP IC50 Assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2366</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Mouse brain cytosol<br />FP-Rh probe<br />Sodium Chloride (Fisher, part 980597)<br />1M Tris, pH 8.0 (Invitrogen, part T-3038)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">ABPP Assay: Purified enzyme (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2371" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2371</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Recombinant PME-1 enzyme<br />FP-Rh probe<br />Sodium Chloride (Fisher, part 980597)<br />1M Tris, pH 8.0 (Invitrogen, part T-3038)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Demethylation (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2363" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2363</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Hela cells<br />HRP-antibody specific to C-terminal demethylated PP2a (Millipore 05-577)<br />Sodium Chloride (Fisher, part 980597)<br />1M Tris, pH 8.0 (Invitrogen, part T-3038)</td></tr><tr><td headers="hd_h_ml136.tu5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Gel Filtration Assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2368" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2368</a>)</td><td headers="hd_h_ml136.tu5_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:middle;">Recombinant PME-1<br />FP-Rh probe<br />Sodium Chloride (Fisher, part 980597)<br />1M Tris, pH 8.0 (Invitrogen, part T-3038)<br />Sephadex G-25 (GE Healthcare, part 17-0851-01)</td></tr></tbody></table></div></div></div><div id="ml136.s11"><h4>iii. Summary of Results</h4><p> Following Primary screening in singlicate (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2130" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2130</a> and <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2269" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2269</a>),
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confirmation of hit activity in triplicate (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2171" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2171</a>), counterscreening by gel-based ABPP in
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proteomes to determine selectivity (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2369" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2369</a> and <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2366</a>), we identified an sulfonyl acrylonitrile
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class of inhibitor for probe development</p><p>CID 44607965 discovered from these efforts represents the first selective covalent PME-1 inhibitor. We anticipate it will be useful to determine the in vivo function of PME-1.</p></div></div><div id="ml136.s12"><h3>c. Probe Optimization</h3><div id="ml136.s13"><h4>i. Description of SAR & chemistry strategy (including structure and data) that led to the probe</h4><p>We speculated that this sulfonyl acrylonitrile core from our hit compound <b>10</b> (<a href="#ml136.s28">Appendix 2</a>, <a class="figpopup" href="/books/NBK50686/table/ml136.t1/?report=objectonly" target="object" rid-figpopup="figml136t1" rid-ob="figobml136t1">Table 1</a>) was covalently reacting with the active site serine of PME-1. An initial round of SAR by purchase and synthetic efforts in the Cravatt lab indeed revealed that this chemical moiety is critical for PME-1 inhibitory activity. Specifically, oxidation of either the olefin (compound <b>20</b>) or the nitrile (compound <b>21</b>) or replacement of the sulfonyl with a ketone (compound <b>32</b>) resulted in completely inactive compounds. Further, the bis-sulfonyl compound <b>19</b> was completely inactive and the bis-nitrile compounds <b>17</b> and <b>18</b> resulted in a 2-fold loss in potency. Once we had determined that the sulfonyl acrylonitrile was important for activity, we investigated the sulfonamide portion of the compound. Our initial round of SAR by purchase (compounds <b>33</b>–<b>48</b>) showed that a wide-range of pyrroles have no activity if not conjugated to a deactivating sulfonamide. Through synthetic efforts, we confirmed and furthered these results by showing that pyrrole itself (compound <b>24</b>) is not active, nor is a benzyl group (compound <b>22</b>) or even the electron-withdrawing para-nitrobenzyl group (compound <b>23</b>). From these two studies, we determined both the sulfonyl acrylonitrile and the sulfonamide are critical for activity.</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml136t1"><a href="/books/NBK50686/table/ml136.t1/?report=objectonly" target="object" title="Table 1" class="img_link icnblk_img figpopup" rid-figpopup="figml136t1" rid-ob="figobml136t1"><img class="small-thumb" src="/books/NBK50686/table/ml136.t1/?report=thumb" src-large="/books/NBK50686/table/ml136.t1/?report=previmg" alt="Table 1. Comparative data on similar compound structures establishing SAR." /></a><div class="icnblk_cntnt"><h4 id="ml136.t1"><a href="/books/NBK50686/table/ml136.t1/?report=objectonly" target="object" rid-ob="figobml136t1">Table 1</a></h4><p class="float-caption no_bottom_margin">Comparative data on similar compound structures establishing SAR. </p></div></div><p>In our next synthetic efforts, we modified the identity of the sulfonamide. We discovered that changing the size of this group (compounds <b>27</b>–<b>30</b>) led to complete loss of activity. Encouragingly, we also discovered that more electron-withdrawing sulfonamides (compounds <b>3</b>–<b>5</b>, <b>7</b>) led to an increase in potency. Specifically, compound <b>3</b> with a meta-cyano group had an IC<sub>50</sub> of 3 μM and was selective for PME-1 relative to other serine hydrolases in the mouse brain soluble proteome. However, the addition of electron-withdrawing nature of the sulfonamide could only increase potency to a point, as the strongly electron-withdawing compounds <b>25</b> and <b>26</b> had no PME-1 inhibitory activity. We next altered the size and electronics of the sulfonyl moiety. Relative to the parent compound <b>10</b>, we discovered that both increasing the size at this position (compound <b>9</b>, IC<sub>50</sub> = 6.8 μM) and making it more electron-withdrawing (compound <b>8</b>, IC<sub>50</sub> = 4.8) led to an increase in potency. At this point, we combined the best modifications on both sides of the molecule, namely meta-cyano substituted sulfonamide and the para-fluoro sulfonyl, and discovered an additive effect resulting in the most potent compound yet (<b>2</b>) with an IC<sub>50</sub> of 640 nM. From this new scaffold we made several minor modifications, including halogen substitution (compounds <b>11</b>, <b>14</b>) and replacement of the meta-cyano (compounds <b>1</b>, <b>12</b>, <b>13</b>, <b>15</b>, <b>16</b>), to discover the probe compound <b>1</b>, which has a meta-nitro group instead of a meta-cyano group. This compound has an IC<sub>50</sub> for PME-1 of 500 nM and was selective for PME-1 relative to other serine hydrolases by >40-fold.</p><p>As a class, the sulfonyl acrylonitrile compounds were highly selective as assessed by their anti-target reactivity by competitive-ABPP. Of the compounds tested at 20µM, only compounds <b>7</b>, <b>10</b>, <b>19</b>, and <b>30</b> showed evidence of anti-target reactivity with a 75kDa serine hydrolase (<a class="figpopup" href="/books/NBK50686/table/ml136.t1/?report=objectonly" target="object" rid-figpopup="figml136t1" rid-ob="figobml136t1">Table 1</a>
|
||
<a href="#ml136.s28">Appendix 2</a>, <a class="figpopup" href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" rid-figpopup="figml136f1" rid-ob="figobml136f1">Figure 1d</a>
|
||
<a href="#ml136.s28">Appendix 2</a>).</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml136f1" co-legend-rid="figlgndml136f1"><a href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" title="Figure 1" class="img_link icnblk_img figpopup" rid-figpopup="figml136f1" rid-ob="figobml136f1"><img class="small-thumb" src="/books/NBK50686/bin/ml136f1.gif" src-large="/books/NBK50686/bin/ml136f1.jpg" alt="Figure 1. Low throughput assays to characterize probe." /></a><div class="icnblk_cntnt" id="figlgndml136f1"><h4 id="ml136.f1"><a href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" rid-ob="figobml136f1">Figure 1</a></h4><p class="float-caption no_bottom_margin">Low throughput assays to characterize probe. <i>A.</i> Selective inhibition of
|
||
PME-1 in the mouse brain soluble proteome as determined by gel-based ABPP. <i>B.</i> Compounds <i>1</i> and <i>2</i> covalently inhibit PME-1, retaining inhibitory activity after gel filtration. <i>C</i>. Hela <a href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" rid-ob="figobml136f1">(more...)</a></p></div></div><p>To date, the lead hit compound <b>10</b> (<a href="#ml136.s28">Appendix 2</a>, <a class="figpopup" href="/books/NBK50686/table/ml136.t1/?report=objectonly" target="object" rid-figpopup="figml136t1" rid-ob="figobml136t1">Table 1</a>) from this inhibitor class has been tested in 152 other bioassays deposited in PubChem and has shown activity in no other systems—further evidence that this compound class does not possess promiscuous activity against other targets.</p></div></div></div><div id="ml136.s14"><h2 id="_ml136_s14_">3. Probe</h2><div id="ml136.s15"><h3>a. Chemical name of probe compound</h3><p>(E)-2-(4-fluorophenylsulfonyl)-3-(1-(3-nitrophenylsulfonyl)-1H-pyrrol-2-yl)acrylonitrile [<a href="/pcsubstance/?term=ML136[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML136</a>]</p></div><div id="ml136.s16"><h3>b. Probe chemical structure including stereochemistry</h3><div id="ml136.fu2" class="figure"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu2.jpg" alt="Image ml136fu2" /></div></div></div><div id="ml136.s17"><h3>c. Structural Verification Information of probe <a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457340" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-87457340</a></h3><ol class="lower-roman"><li class="half_rhythm"><div>1H NMR (500 MHz, CDCl<sub>3</sub>) δ 8.78 (d, J = 0.6 Hz, 1H), 8.68 (dd, J = 2.0, 2.0 Hz, 1H), 8.58-8.53 (m, 1H), 8.29-8.26 (m, 1H), 8.03 (dd, J =8.7, 4.9 Hz, 2H), 7.87 (dd, J =8.1, 8.1 Hz, 1H), 7.77-7.75 (m, 1H), 7.67-7.64 (m, 1H), 7.30 (dd, J = 8.4, 8.4 Hz, 2H), 6.59 (dd, J = 3.9, 3.3 HZ, 1H), purity >95%</div></li><li class="half_rhythm"><div>high-res MS analysis (ESI-TOF): M-H<sup>−</sup> expected: 460.0079, M-H<sup>−</sup> observed: 460.0096</div></li></ol></div><div id="ml136.s18"><h3>d. PubChem CID (corresponding to the SID)</h3><p>CID 44607965</p></div><div id="ml136.s19"><h3>e. Availability from a Vendor</h3><p>Not available</p></div><div id="ml136.s20"><h3>f. MLS#'s of probe molecule and five related samples that were submitted to the SMR collection</h3><p>MLS002699139 (<a class="figpopup" href="/books/NBK50686/table/ml136.tu2/?report=objectonly" target="object" rid-figpopup="figml136tu2" rid-ob="figobml136tu2">see table p.2</a>)</p></div><div id="ml136.s21"><h3>g. Mode of action for biological activity of probe</h3><p>We have shown that CID 44607965 (referred as compound <b>1</b> in <a href="#ml136.s28">Appendix 2</a>) forms a covalent adduct with PME-1 by demonstrating that blockade of FP-rhodamine labeling is not reversed by gel filtration (<a href="#ml136.s28">Appendix 2</a>, <a class="figpopup" href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" rid-figpopup="figml136f1" rid-ob="figobml136f1">Figure 1B</a>). Even though <b>1</b> contains a reactive chemical moiety, this compound selectively inhibits PME-1 in the mouse brain soluble proteome (<a href="#ml136.s28">Appendix 2</a>, <a class="figpopup" href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" rid-figpopup="figml136f1" rid-ob="figobml136f1">Figure 1A</a>). Further, we showed that the probe inhibited the ability of PME-1 to demethylate PP2A in HeLa cells as evidenced by a decrease in demethylated PP2A after incubation of cells with compound (<a href="#ml136.s28">Appendix 2</a>, <a class="figpopup" href="/books/NBK50686/figure/ml136.f1/?report=objectonly" target="object" rid-figpopup="figml136f1" rid-ob="figobml136f1">Figure 1C</a>).</p></div><div id="ml136.s22"><h3>h. Detailed synthetic pathway</h3><div id="ml136.fu3" class="figure"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu3.jpg" alt="Image ml136fu3" /></div></div><p><b>(<i>E</i>)-2-(4-fluorophenylsulfonyl)-3-(1<i>H</i>-pyrrol-2-yl)acrylonitrile.</b> Powdered 4A
|
||
molecular sieves were charged into a 25mL round bottom flask and flame dried under vacuum. After
|
||
replacing the atmosphere with nitrogen, ethanol (12.5mL) was added, followed by
|
||
pyrrole-2-carboxaldehyde (200mg, 2.1mmol, 1eq) the sulfonyl acetonitrile (419mg, 2.1mmol, 1eq), and
|
||
triethylamine (1.25mL, 9.0mmol, 4.3eq). The reaction was refluxed for 4 hr, cooled to room
|
||
temperature and then quenched with water (5mL). The aqueous layer was acidified with 1N HCl (2mL)
|
||
and the product extracted into DCM (3 x 20mL). The combined organic layers were then washed with
|
||
water (10mL), brine (10mL), dried over MgS0<sub>4</sub> and concentrated <i>in vacuo</i>.
|
||
Flash chromatography (SiO<sub>2</sub>, 10-50% ethyl acetate/hexanes) afforded the free pyrrole
|
||
product (498 mg, 1.81mmol, 86%).</p><p><b>(<i>E</i>)-2-(4-fluorophenylsulfonyl)-3-(1-(3-nitrophyeylsulfonyl)-1<i>H</i>-pyrrol-2-yl)acrylonitrile
|
||
(PROBE AMZ30).</b> Under nitrogen atmosphere, a 5mL flame-dried round bottom flask was charged
|
||
with the free pyrrole (10mg, 0.036mmol, 1eq). The solid was diluted with dimethylformamide (0.580mL,
|
||
0.05M). Triethylamine (21μL, 0.145mmol, 5eq) was then added, followed by
|
||
3-nitrobenzenesulfonyl chloride (9.6mg, 0.043mmol, 1.2eq). The reaction was allowed to stir
|
||
overnight at which point the solvent was removed <i>in vacuo</i>. Purification by
|
||
preparatory TLC (SiO<sub>2</sub>, 1000μm, 50% ethyl acetate/hexanes) afforded the product
|
||
(14.1mg, 0.030mmol, 84%).</p></div><div id="ml136.s23"><h3>i. Summary of probe properties (solubility, absorbance/fluorescence, reactivity, toxicity, etc.)</h3><p>ADMET BBB, undefined; ADMET BBB level, undefined; ADMET absorption level, 2; ADMET solubility, −5.031; ADMET solubility level, low.</p><p>Solubility of the probe in PBS (137 mM NaCl, 2.7 mM KCl, 10 mM sodium phosphate dibasic, 2 mM potassium phosphate monobasic, pH 7.4) at room temperature was determined to be 28.4 µM. The probe has a half-life of >48 hours in PBS at room temperature (tested at 10 µM, <a class="figpopup" href="/books/NBK50686/figure/ml136.f3/?report=objectonly" target="object" rid-figpopup="figml136f3" rid-ob="figobml136f3">Figure 3</a>
|
||
<a href="#ml136.s28">Appendix 2</a>).</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml136f3" co-legend-rid="figlgndml136f3"><a href="/books/NBK50686/figure/ml136.f3/?report=objectonly" target="object" title="Figure 3" class="img_link icnblk_img figpopup" rid-figpopup="figml136f3" rid-ob="figobml136f3"><img class="small-thumb" src="/books/NBK50686/bin/ml136f3.gif" src-large="/books/NBK50686/bin/ml136f3.jpg" alt="Figure 3. Stability of ML136 (Compound 1) in PBS indicates a half-life of >48 hours." /></a><div class="icnblk_cntnt" id="figlgndml136f3"><h4 id="ml136.f3"><a href="/books/NBK50686/figure/ml136.f3/?report=objectonly" target="object" rid-ob="figobml136f3">Figure 3</a></h4><p class="float-caption no_bottom_margin">Stability of ML136 (Compound 1) in PBS indicates a half-life of >48 hours. </p></div></div><p>The probe compound showed no reactivity with glutathione (100 µM), indicating that it is not generally cysteine reactive, but rather has a tempered electrophilicity and specific structural elements that direct reactivity towards GSTO1. An irreversible probe has some distinct advantages over reversible analogs. Targets can be readily characterized by methods such as mass spectrometry and click chemistry-ABPP, required dosing is often lower, irreversible compounds are not as sensitive to pharmacokinetic parameters, and administration can induce long-lasting inhibition [<a class="bk_pop" href="#ml136.r11">11</a>]. In the case of the EGFR inhibitor PD 0169414, its irreversibility and high selectivity were credited with producing prolonged inhibition of the target, alleviating concerns over short plasma half-lives and reducing the need for high peak plasma levels, thus minimizing potential nonspecific toxic effects [<a class="bk_pop" href="#ml136.r12">12</a>].</p><p>Indeed, over a third of enzymatic drug targets are irreversibly inhibited by currently marketed drugs [<a class="bk_pop" href="#ml136.r13">13</a>]. Examples of covalent enzyme-inhibitor pairs include serine type D-Ala-D-Ala carboxypeptidase, which is covalently modified by all beta-lactam antibiotics, acetylcholinesterase, whose active site serine undergoes covalent modification by pyridostigmine, prostaglandin-endoperoxide synthase, which is the target of the ubiquitously prescribed aspirin, aromatase, which is irreversibly modified by exemestane, monoamine oxidase, which is covalently modified by L-deprenyl, thymidylate synthase, which is covalently modified by floxuridine, H+/K+ ATPase, which undergoes covalent modification by omaprazole, esmoprazole, and lanoprazole, and triacylglycerol lipase, whose serine nucleophile is targeted by orlistat [<a class="bk_pop" href="#ml136.r13">13</a>].</p></div><div id="ml136.s24"><h3>j. Probe properties</h3><div id="ml136.tu6" class="table"><h3><span class="title">Properties Computed from Structure</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu6/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu6_lrgtbl__"><table class="no_margin"><tbody><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">PubChem CID</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">CID 44607965</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">PubChem SID</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457340" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-87457340</a></td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">IUPAC Name</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">(E)-2-(4-fluorophenylsulfonyl)-3-(1-(3-nitrophenylsulfonyl)-1H-pyrrol-2-yl)acrylonitrile</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">MLS</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">MLS002699139</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">MF</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">C<sub>19</sub>H<sub>12</sub>FN<sub>3</sub>O<sub>6</sub>S<sub>2</sub></td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">MW</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">461.44</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Formal Charge</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">H Acceptor</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">6</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">H Donor</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Atom Count</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">43</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Rotatable Bonds</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">6</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Rings</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">3</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Stereoatoms</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">1</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">AlogP</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">3.825</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">logD</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">3.825</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Polar surface area</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">155.21</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Aqueous solubility<sup><a class="bk_pop" href="#ml136.tfn3">a</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">28.4 µM</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Aqueous stability<sup><a class="bk_pop" href="#ml136.tfn3">a</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">half-life >48 hours</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Reactivity with Glutathione<sup><a class="bk_pop" href="#ml136.tfn3">a</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">None</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Mechanism of Action</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Irreversible (covalent) inhibitor of PME-1</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">ADMET BBB<sup><a class="bk_pop" href="#ml136.tfn4">b</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Undefined</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">ADMET BBB level<sup><a class="bk_pop" href="#ml136.tfn5">c</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Undefined</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">ADMET absorption level<sup><a class="bk_pop" href="#ml136.tfn6">d</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">2 (Low)</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">ADMET solubility<sup><a class="bk_pop" href="#ml136.tfn7">e</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">−5.031</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">ADMET solubility level<sup><a class="bk_pop" href="#ml136.tfn7">e</a></sup></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Low</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Vendor</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">None</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Vendor Catalog Number</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">None</td></tr></tbody></table></div><div><div><dl class="temp-labeled-list small"><dt>a</dt><dd><div id="ml136.tfn3"><p class="no_margin">Determined according to NIH guidelines.</p></div></dd><dt>b</dt><dd><div id="ml136.tfn4"><p class="no_margin">ADMET_BBB: Log of Brain/Blood partition coefficient (LogBB) [<a class="bk_pop" href="#ml136.r14">14</a>].</p></div></dd><dt>c</dt><dd><div id="ml136.tfn5"><p class="no_margin">ADMET_BBB_Level: Ranking of the LogBB values into one of the following levels: 0: Very High, 1: High, 2: Medium, 3: Low, 4: Undefined (molecule is outside the confidence area of the regression model) [<a class="bk_pop" href="#ml136.r14">14</a>, <a class="bk_pop" href="#ml136.r15">15</a>].</p></div></dd><dt>d</dt><dd><div id="ml136.tfn6"><p class="no_margin">ADMET Passive Intestinal Absorption properties. Ranking of the molecule into one of the following levels: 0: Good, 1: Moderate, 2: Poor, 3: Very Poor [<a class="bk_pop" href="#ml136.r14">14</a>, <a class="bk_pop" href="#ml136.r15">15</a>].</p></div></dd><dt>e</dt><dd><div id="ml136.tfn7"><p class="no_margin">ADMET_Solubility: Log of the water solubility at 25 degrees, LogSw, in mol/L [<a class="bk_pop" href="#ml136.r14">14</a>, <a class="bk_pop" href="#ml136.r15">15</a>].</p></div></dd></dl></div></div></div></div><div id="ml136.s25"><h3>k. Dose Response Curve for Probe</h3><p>Below is the IC50 Curve for Probe Compound as determined by gel-based competitive-ABPP with FP-Rh. Calculated IC50 = 0.50 µM.</p><div id="ml136.fu4" class="figure"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu4.jpg" alt="Image ml136fu4" /></div></div></div></div><div id="ml136.s26"><h2 id="_ml136_s26_">4. Appendices</h2><div id="ml136.s27"><h3>Appendix 1. PME-1 Inhibitors SAR Table</h3><div id="ml136.tu7" class="table"><h3><span class="title">PME-1 Inhibitors SAR Table</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.tu7/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.tu7_lrgtbl__"><table class="no_margin"><thead><tr><th id="hd_h_ml136.tu7_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></th><th id="hd_h_ml136.tu7_1_1_1_9" colspan="5" rowspan="1" style="text-align:center;vertical-align:middle;">Probe Development Assays</th></tr><tr><th headers="hd_h_ml136.tu7_1_1_1_1" id="hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Compound</th><th headers="hd_h_ml136.tu7_1_1_1_2" id="hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Scripps ID</th><th headers="hd_h_ml136.tu7_1_1_1_3" id="hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Structure</th><th headers="hd_h_ml136.tu7_1_1_1_4" id="hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">CID</th><th headers="hd_h_ml136.tu7_1_1_1_5" id="hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SID</th><th headers="hd_h_ml136.tu7_1_1_1_6" id="hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS ID</th><th headers="hd_h_ml136.tu7_1_1_1_7" id="hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Vendor</th><th headers="hd_h_ml136.tu7_1_1_1_8" id="hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Vendor Catalog ID</th><th headers="hd_h_ml136.tu7_1_1_1_9" id="hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1 ABPP % INH (20μM) [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2369" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2369</a>]</th><th headers="hd_h_ml136.tu7_1_1_1_9" id="hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PME-1 ABPP IC50 (μM) [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2366" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2366</a>]</th><th headers="hd_h_ml136.tu7_1_1_1_9" id="hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Gel Filtration Assay [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2368" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2368</a>]</th><th headers="hd_h_ml136.tu7_1_1_1_9" id="hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Cytotoxicity Assay (CC50) (nM) [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2365" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2365</a>]</th><th headers="hd_h_ml136.tu7_1_1_1_9" id="hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Demethylation Assay % INH (20 μM) [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2363" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2363</a>]</th></tr></thead><tbody><tr><td headers="hd_h_ml136.tu7_1_1_1_1 hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>PROBE</b></td><td headers="hd_h_ml136.tu7_1_1_1_2 hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SR-02000000252-1</td><td headers="hd_h_ml136.tu7_1_1_1_3 hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu5.jpg" alt="Image ml136fu5.jpg" /></div></td><td headers="hd_h_ml136.tu7_1_1_1_4 hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607965</td><td headers="hd_h_ml136.tu7_1_1_1_5 hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">87457340</td><td headers="hd_h_ml136.tu7_1_1_1_6 hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699139</td><td headers="hd_h_ml136.tu7_1_1_1_7 hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_8 hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">90</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.5</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Irreversible</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">100000</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">98</td></tr><tr><td headers="hd_h_ml136.tu7_1_1_1_1 hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 1</b></td><td headers="hd_h_ml136.tu7_1_1_1_2 hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SR-02000000253-1 SR-</td><td headers="hd_h_ml136.tu7_1_1_1_3 hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu6.jpg" alt="Image ml136fu6.jpg" /></div></td><td headers="hd_h_ml136.tu7_1_1_1_4 hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607949</td><td headers="hd_h_ml136.tu7_1_1_1_5 hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">87457341</td><td headers="hd_h_ml136.tu7_1_1_1_6 hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699140</td><td headers="hd_h_ml136.tu7_1_1_1_7 hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_8 hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn8">*</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.64</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Irreversible</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">100000</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">95</td></tr><tr><td headers="hd_h_ml136.tu7_1_1_1_1 hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 2</b></td><td headers="hd_h_ml136.tu7_1_1_1_2 hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">02000000255-1</td><td headers="hd_h_ml136.tu7_1_1_1_3 hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu7.jpg" alt="Image ml136fu7.jpg" /></div></td><td headers="hd_h_ml136.tu7_1_1_1_4 hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607954</td><td headers="hd_h_ml136.tu7_1_1_1_5 hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">87457343</td><td headers="hd_h_ml136.tu7_1_1_1_6 hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699141</td><td headers="hd_h_ml136.tu7_1_1_1_7 hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_8 hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn8">*</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">3.0</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn9">†</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn10">‡</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn11">§</a></sup></td></tr><tr><td headers="hd_h_ml136.tu7_1_1_1_1 hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 3</b></td><td headers="hd_h_ml136.tu7_1_1_1_2 hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SR-02000000259-1</td><td headers="hd_h_ml136.tu7_1_1_1_3 hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu8.jpg" alt="Image ml136fu8.jpg" /></div></td><td headers="hd_h_ml136.tu7_1_1_1_4 hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607974</td><td headers="hd_h_ml136.tu7_1_1_1_5 hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">87457347</td><td headers="hd_h_ml136.tu7_1_1_1_6 hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699142</td><td headers="hd_h_ml136.tu7_1_1_1_7 hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_8 hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn8">*</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">4.8</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn9">†</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn10">‡</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn11">§</a></sup></td></tr><tr><td headers="hd_h_ml136.tu7_1_1_1_1 hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 4</b></td><td headers="hd_h_ml136.tu7_1_1_1_2 hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SR-02000000260-1</td><td headers="hd_h_ml136.tu7_1_1_1_3 hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu9.jpg" alt="Image ml136fu9.jpg" /></div></td><td headers="hd_h_ml136.tu7_1_1_1_4 hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">44607969</td><td headers="hd_h_ml136.tu7_1_1_1_5 hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">87457348</td><td headers="hd_h_ml136.tu7_1_1_1_6 hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699143</td><td headers="hd_h_ml136.tu7_1_1_1_7 hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_8 hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn8">*</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">6.8</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn9">†</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn10">‡</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn11">§</a></sup></td></tr><tr><td headers="hd_h_ml136.tu7_1_1_1_1 hd_h_ml136.tu7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><b>Analog 5</b></td><td headers="hd_h_ml136.tu7_1_1_1_2 hd_h_ml136.tu7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SR-01000639085-3</td><td headers="hd_h_ml136.tu7_1_1_1_3 hd_h_ml136.tu7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><div class="graphic"><img src="/books/NBK50686/bin/ml136fu10.jpg" alt="Image ml136fu10.jpg" /></div></td><td headers="hd_h_ml136.tu7_1_1_1_4 hd_h_ml136.tu7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5703330</td><td headers="hd_h_ml136.tu7_1_1_1_5 hd_h_ml136.tu7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">87457336</td><td headers="hd_h_ml136.tu7_1_1_1_6 hd_h_ml136.tu7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699144</td><td headers="hd_h_ml136.tu7_1_1_1_7 hd_h_ml136.tu7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Maybridge</td><td headers="hd_h_ml136.tu7_1_1_1_8 hd_h_ml136.tu7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="/protein/1094266001/?report=GenPept" class="bk_tag" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=genpept">SEW03242</a></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below <sup><a class="bk_pop" href="#ml136.tfn8">*</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">10.8</td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn9">†</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn10">‡</a></sup></td><td headers="hd_h_ml136.tu7_1_1_1_9 hd_h_ml136.tu7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">See below<sup><a class="bk_pop" href="#ml136.tfn11">§</a></sup></td></tr></tbody></table></div><div><div><dl class="temp-labeled-list small"><dt>*</dt><dd><div id="ml136.tfn8"><p class="no_margin">Only the more precise IC<sub>50</sub> value is reported.</p></div></dd><dt>†</dt><dd><div id="ml136.tfn9"><p class="no_margin">These compounds were not tested in this assay because this is a mechanistic assay, and these related compounds are expected to all interact in the same covalent manner with PME-1.</p></div></dd><dt>‡</dt><dd><div id="ml136.tfn10"><p class="no_margin">Not Tested.</p></div></dd><dt>§</dt><dd><div id="ml136.tfn11"><p class="no_margin">Not Tested.</p></div></dd></dl></div></div></div></div><div id="ml136.s28"><h3>Appendix 2. Assay Provider/Probe Development Assays</h3><div id="ml136.f2" class="figure bk_fig"><div class="graphic"><img src="/books/NBK50686/bin/ml136f2.jpg" alt="Figure 2. Cytotoxicity of PME-1 inhibitors against HEK 293T cells after 48h of treatment as determined by the CellTitre assay (Promega)." /></div><h3><span class="label">Figure 2</span><span class="title">Cytotoxicity of PME-1 inhibitors against HEK 293T cells after 48h of treatment as determined
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by the CellTitre assay (Promega)</span></h3><div class="caption"><p>Both compounds <b>1</b> (AMZ30) and
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<b>2</b> (DAB8) are cytotoxic only at very high concentrations (CC<sub>50</sub>~100 μM), well above the concentrations used to generate effects on PP2A methylation (<20 μM)</p></div></div></div><div id="ml136.pmidlist"><h3>PMID List</h3><div id="ml136.pmidtable" class="table"><h3><span class="title">PMID List</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK50686/table/ml136.pmidtable/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml136.pmidtable_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Ref</th><th id="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">PMID</th></tr></thead><tbody><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">9727084</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11171037</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">3</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">15661531</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">4</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">8650216</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11060018</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">6</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">19293187</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">7</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">18596935</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">8</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">12149457</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">9</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">12740587</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">10</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">19329999</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">20640225</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">12</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">10663641</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">13</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">15823014</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">14</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11052792</td></tr><tr><td headers="hd_h_ml136.pmidtable_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">15</td><td headers="hd_h_ml136.pmidtable_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11922948</td></tr></tbody></table></div></div></div></div><div id="ml136.s29"><h2 id="_ml136_s29_">5. References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="ml136.r1">Oliver CJ, Shenolikar S. Physiologic importance of protein phosphatase inhibitors. <span><span class="ref-journal">Front. Biosci. </span>1998;<span class="ref-vol"><strong>3</strong></span>:D961–72.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/9727084" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 9727084</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="ml136.r2">Janssens V, Goris J. Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. <span><span class="ref-journal">Biochem. J. </span>2001;<span class="ref-vol"><strong>353</strong></span>(Pt 3):417–39.</span> [<a href="/pmc/articles/PMC1221586/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC1221586</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/11171037" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11171037</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="ml136.r3">Janssens V, Goris J, Van Hoof C. PP2A: the expected tumor suppressor. <span><span class="ref-journal">Curr. Opin. Genet. Dev. </span>2005;<span class="ref-vol"><strong>15</strong></span>(1):34–41.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15661531" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15661531</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="ml136.r4">Lee J, et al. A specific protein carboxyl methylesterase that demethylates phosphoprotein phosphatase 2A in bovine brain. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>1996;<span class="ref-vol"><strong>93</strong></span>(12):6043–7.</span> [<a href="/pmc/articles/PMC39185/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC39185</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/8650216" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 8650216</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="ml136.r5">Wu J, et al. Carboxyl methylation of the phosphoprotein phosphatase 2A catalytic subunit promotes its functional association with regulatory subunits in vivo. <span><span class="ref-journal">EMBO J. </span>2000;<span class="ref-vol"><strong>19</strong></span>(21):5672–81.</span> [<a href="/pmc/articles/PMC305778/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC305778</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/11060018" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11060018</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="ml136.r6">Puustinen P, et al. PME-1 protects extracellular signal-regulated kinase pathway activity from protein phosphatase 2A-mediated inactivation in human malignant glioma. <span><span class="ref-journal">Cancer Res. </span>2009;<span class="ref-vol"><strong>69</strong></span>(7):2870–7.</span> [<a href="/pmc/articles/PMC2810347/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2810347</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/19293187" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 19293187</span></a>]</div></dd><dt>7.</dt><dd><div class="bk_ref" id="ml136.r7">Ortega-Gutierrez S, et al. Targeted disruption of the PME-1 gene causes loss of demethylated PP2A and perinatal lethality in mice. <span><span class="ref-journal">PLoS One. </span>2008;<span class="ref-vol"><strong>3</strong></span>(7):e2486.</span> [<a href="/pmc/articles/PMC2438471/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2438471</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/18596935" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18596935</span></a>]</div></dd><dt>8.</dt><dd><div class="bk_ref" id="ml136.r8">Jessani N, et al. Enzyme activity profiles of the secreted and membrane proteome that depict cancer cell invasiveness. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>2002;<span class="ref-vol"><strong>99</strong></span>(16):10335–40.</span> [<a href="/pmc/articles/PMC124915/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC124915</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/12149457" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12149457</span></a>]</div></dd><dt>9.</dt><dd><div class="bk_ref" id="ml136.r9">Leung D, et al. Discovering potent and selective reversible inhibitors of enzymes in complex proteomes. <span><span class="ref-journal">Nat Biotechnol. </span>2003;<span class="ref-vol"><strong>21</strong></span>(6):687–91.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12740587" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12740587</span></a>]</div></dd><dt>10.</dt><dd><div class="bk_ref" id="ml136.r10">Bachovchin DA, et al. Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes. <span><span class="ref-journal">Nat Biotechnol. </span>2009;<span class="ref-vol"><strong>27</strong></span>(4):387–94.</span> [<a href="/pmc/articles/PMC2709489/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2709489</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/19329999" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 19329999</span></a>]</div></dd><dt>11.</dt><dd><div class="bk_ref" id="ml136.r11">Johnson DS, Weerapana E, Cravatt BF. Strategies for discovering and derisking covalent, irreversible enzyme inhibitors. <span><span class="ref-journal">Future Med Chem. </span>2010;<span class="ref-vol"><strong>2</strong></span>(6):949–964.</span> [<a href="/pmc/articles/PMC2904065/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2904065</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/20640225" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 20640225</span></a>]</div></dd><dt>12.</dt><dd><div class="bk_ref" id="ml136.r12">Vincent PW, et al. Anticancer efficacy of the irreversible EGFr tyrosine kinase inhibitor PD 0169414 against human tumor xenografts. <span><span class="ref-journal">Cancer Chemother Pharmacol. </span>2000;<span class="ref-vol"><strong>45</strong></span>(3):231–8.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10663641" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10663641</span></a>]</div></dd><dt>13.</dt><dd><div class="bk_ref" id="ml136.r13">Robertson JG. Mechanistic basis of enzyme-targeted drugs. <span><span class="ref-journal">Biochemistry. </span>2005;<span class="ref-vol"><strong>44</strong></span>(15):5561–71.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15823014" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15823014</span></a>]</div></dd><dt>14.</dt><dd><div class="bk_ref" id="ml136.r14">Egan WJ, Merz KM Jr, Baldwin JJ. Prediction of drug absorption using multivariate statistics. <span><span class="ref-journal">J Med Chem. </span>2000;<span class="ref-vol"><strong>43</strong></span>(21):3867–77.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11052792" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11052792</span></a>]</div></dd><dt>15.</dt><dd><div class="bk_ref" id="ml136.r15">Egan WJ, Lauri G. Prediction of intestinal permeability. <span><span class="ref-journal">Adv Drug Deliv Rev. </span>2002;<span class="ref-vol"><strong>54</strong></span>(3):273–89.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11922948" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11922948</span></a>]</div></dd></dl></div><div id="bk_toc_contnr"></div></div></div>
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<div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Views</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PDF_download" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="/books/NBK50686/?report=reader">PubReader</a></li><li><a href="/books/NBK50686/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK50686" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK50686" style="display:none" title="Cite this Page"><div class="bk_tt">Bachovchin DA, Speers AE, Zuhl AM, et al. Probe Report for PME-1 Inhibitors. 2010 Feb 26 [Updated 2010 Oct 20]. In: Probe Reports from the NIH Molecular Libraries Program [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2010-. <span class="bk_cite_avail"></span></div></div></li><li><a href="/books/NBK50686/pdf/Bookshelf_NBK50686.pdf">PDF version of this page</a> (766K)</li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>In this Page</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="page-toc" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="#ml136.s2" ref="log$=inpage&link_id=inpage">Probe Structure & Characteristics</a></li><li><a href="#ml136.s3" ref="log$=inpage&link_id=inpage">Probe Selection</a></li><li><a href="#ml136.s4" ref="log$=inpage&link_id=inpage">Recommendations for the Scientific Use of This Probe</a></li><li><a href="#ml136.s5" ref="log$=inpage&link_id=inpage">Scientific Rationale for Project</a></li><li><a href="#ml136.s6" ref="log$=inpage&link_id=inpage">Project Description</a></li><li><a href="#ml136.s14" ref="log$=inpage&link_id=inpage">Probe</a></li><li><a href="#ml136.s26" ref="log$=inpage&link_id=inpage">Appendices</a></li><li><a href="#ml136.s29" ref="log$=inpage&link_id=inpage">References</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pmc&DbFrom=books&Cmd=Link&LinkName=books_pmc_refs&IdsFromResult=2376368" ref="log$=recordlinks">PMC</a><div class="brieflinkpop offscreen_noflow">PubMed Central citations</div></li><li 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inhibitors of protein phosphatase methylesterase-1.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Bachovchin DA, Zuhl AM, Speers AE, Wolfe MR, Weerapana E, Brown SJ, Rosen H, Cravatt BF. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">J Med Chem. 2011 Jul 28; 54(14):5229-36. Epub 2011 Jun 30.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/24928782" ref="ordinalpos=1&linkpos=3&log$=relatedarticles&logdbfrom=pubmed">PME-1 modulates protein phosphatase 2A activity to promote the malignant phenotype of endometrial cancer cells.</a><span class="source">[Cancer Res. 2014]</span><div class="brieflinkpop offscreen_noflow">PME-1 modulates protein phosphatase 2A activity to promote the malignant phenotype of endometrial cancer cells.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Wandzioch E, Pusey M, Werda A, Bail S, Bhaskar A, Nestor M, Yang JJ, Rice LM. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Cancer Res. 2014 Aug 15; 74(16):4295-305. Epub 2014 Jun 13.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21398589" ref="ordinalpos=1&linkpos=4&log$=relatedarticles&logdbfrom=pubmed">Academic cross-fertilization by public screening yields a remarkable class of protein phosphatase methylesterase-1 inhibitors.</a><span class="source">[Proc Natl Acad Sci U S A. 2011]</span><div class="brieflinkpop offscreen_noflow">Academic cross-fertilization by public screening yields a remarkable class of protein phosphatase methylesterase-1 inhibitors.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Bachovchin DA, Mohr JT, Speers AE, Wang C, Berlin JM, Spicer TP, Fernandez-Vega V, Chase P, Hodder PS, Schürer SC, et al. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Proc Natl Acad Sci U S A. 2011 Apr 26; 108(17):6811-6. Epub 2011 Mar 11.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/23762952" ref="ordinalpos=1&linkpos=5&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Probe Development Efforts to Identify Novel Inhibitors of ABHD10.</a><span class="source">[Probe Reports from the NIH Mol...]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Probe Development Efforts to Identify Novel Inhibitors of ABHD10.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Zuhl AM, Mohr JT, Speers AE, Bachovchin DA, Berlin JM, Spicer T, Fernandez-Vega V, Brown SJ, Ferguson J, Fu GC, et al. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Probe Reports from the NIH Molecular Libraries Program. 2010</em></div></div></li></ul><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed_reviews&uid=21433379" ref="ordinalpos=1&log$=relatedreviews_seeall&logdbfrom=pubmed">See reviews...</a><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=21433379" ref="ordinalpos=1&log$=relatedarticles_seeall&logdbfrom=pubmed">See all...</a></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Recent Activity</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="recent_activity" id="Shutter"></a></div><div class="portlet_content"><div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" id="HTDisplay" class=""><div class="action"><a href="javascript:historyDisplayState('ClearHT')">Clear</a><a href="javascript:historyDisplayState('HTOff')" class="HTOn">Turn Off</a><a href="javascript:historyDisplayState('HTOn')" class="HTOff">Turn On</a></div><ul id="activity"><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=1" href="/portal/utils/pageresolver.fcgi?recordid=67d66c0567c23b31e0b36ef0">Probe Report for PME-1 Inhibitors - 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