Purification and characterization of recombinant microsomal prostaglandin E synthase-1
- PMID: 12460774
- DOI: 10.1016/s1046-5928(02)00566-1
Purification and characterization of recombinant microsomal prostaglandin E synthase-1
Abstract
Recombinant human microsomal prostaglandin E(2) synthase-1 (mPGES-1) was expressed in a baculovirus-Sf9 cell system. The mPGES-1 was solubilized from Sf9 cell membranes with diheptanoylphosphatidylcholine and purified in the presence of octylglucoside using hydroxyapatite column chromatography. The K(m) values of the substrates PGH(2) and GSH were 14 microM and 0.75 mM, respectively, with the purified enzyme. The specific activity (4 micromol/min/mg) was increased 3-5-fold by non-ionic and zwitterionic detergents. Kinetic analysis showed that dodecylmaltoside increases V(max) but does not affect the K(m) values of either substrate. Several other thiol-containing compounds were tested as glutathione replacements, none of which yielded detectable enzyme activity. During enzyme catalysis, glutathione was not oxidized and therefore can be considered an enzyme cofactor. No glutathione transferase or peroxidase activity could be determined with a range of potential substrates. The results show that purified mPGES-1 has a specific activity similar to Cox-2, consistent with its postulated role in Cox-2 mediated PGE(2) formation.
Similar articles
-
Human microsomal prostaglandin E synthase-1: purification, functional characterization, and projection structure determination.J Biol Chem. 2003 Jun 20;278(25):22199-209. doi: 10.1074/jbc.M303227200. Epub 2003 Apr 2. J Biol Chem. 2003. PMID: 12672824
-
Purification and characterization of membrane-bound prostaglandin E synthase from bovine heart.Biochim Biophys Acta. 1999 Aug 18;1439(3):406-14. doi: 10.1016/s1388-1981(99)00084-0. Biochim Biophys Acta. 1999. PMID: 10446427
-
Purification and characterization of recombinant human prostacyclin synthase.J Biochem. 2004 Apr;135(4):455-63. doi: 10.1093/jb/mvh059. J Biochem. 2004. PMID: 15115769
-
Membrane prostaglandin E synthase-1: a novel therapeutic target.Pharmacol Rev. 2007 Sep;59(3):207-24. doi: 10.1124/pr.59.3.1. Pharmacol Rev. 2007. PMID: 17878511 Review.
-
Microsomal prostaglandin E synthase-1 and 5-lipoxygenase: potential drug targets in cancer.J Intern Med. 2010 Jul;268(1):5-14. doi: 10.1111/j.1365-2796.2010.02246.x. Epub 2010 Apr 28. J Intern Med. 2010. PMID: 20497297 Review.
Cited by
-
Enzymes of the cyclooxygenase pathways of prostanoid biosynthesis.Chem Rev. 2011 Oct 12;111(10):5821-65. doi: 10.1021/cr2002992. Epub 2011 Sep 27. Chem Rev. 2011. PMID: 21942677 Free PMC article. Review. No abstract available.
-
Crystal structure of microsomal prostaglandin E2 synthase provides insight into diversity in the MAPEG superfamily.Proc Natl Acad Sci U S A. 2013 Mar 5;110(10):3806-11. doi: 10.1073/pnas.1218504110. Epub 2013 Feb 19. Proc Natl Acad Sci U S A. 2013. PMID: 23431194 Free PMC article.
-
Competitive enzymatic interactions determine the relative amounts of prostaglandins E2 and D2.J Pharmacol Exp Ther. 2011 Nov;339(2):716-25. doi: 10.1124/jpet.111.185405. Epub 2011 Aug 24. J Pharmacol Exp Ther. 2011. PMID: 21865441 Free PMC article.
-
Prostaglandin E2 and Receptors: Insight Into Tumorigenesis, Tumor Progression, and Treatment of Hepatocellular Carcinoma.Front Cell Dev Biol. 2022 Mar 10;10:834859. doi: 10.3389/fcell.2022.834859. eCollection 2022. Front Cell Dev Biol. 2022. PMID: 35356289 Free PMC article. Review.
-
Omega-3 fatty acids cause dramatic changes in TLR4 and purinergic eicosanoid signaling.Proc Natl Acad Sci U S A. 2012 May 29;109(22):8517-22. doi: 10.1073/pnas.1200189109. Epub 2012 May 14. Proc Natl Acad Sci U S A. 2012. PMID: 22586114 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases
Research Materials
