Open AccessHTRF-Based Assay for Microsomal Prostaglandin E2 Synthase-1 Activity
Author(s) -
Eric R. Goedken,
Andrew Gag,
Gary T. Overmeyer,
Junjian Liu,
Richard A. Petrillo,
Andrew Burchat,
Medha J. Tomlinson
Publication year - 2008
Publication title -
slas discovery
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 0.907
H-Index - 75
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/1087057108321145
Subject(s) - microsome , prostaglandin e2 , enzyme , prostaglandin , chemistry , enzyme assay , atp synthase , in vitro , potency , substrate (aquarium) , prostaglandin e , biochemistry , bioassay , prostaglandin h2 , in vivo , biology , microbiology and biotechnology , arachidonic acid , ecology , genetics , endocrinology
Microsomal prostaglandin E2 synthase-1 (mPGES-1) catalyzes the formation of prostaglandin E2 (PGE2) from the endoperoxide prostaglandin H2 (PGH2). Expression of this enzyme is induced during the inflammatory response, and mouse knockout experiments suggest it may be an attractive target for antiarthritic therapies. Assaying the activity of this enzyme in vitro is challenging because of the unstable nature of the PGH2 substrate. Here, the authors present an mPGES-1 activity assay suitable for characterization of enzyme preparations and for determining the potency of inhibitor compounds. This plate-based competition assay uses homogenous time-resolved fluorescence to measure PGE2 produced by the enzyme. The assay is insensitive to DMSO concentration up to 10% and does not require extensive washes after the initial enzyme reaction is concluded, making it a simple and convenient way to assess mPGES-1 inhibition.