Cyclooxygenase (COX, also called Prostaglandin H Synthase or PGHS) is a bifunctional enzyme exhibiting both COX and peroxidase activities. The COX component converts arachidonic acid to a hydroperoxy endoperoxide (PGG2), and the peroxidase component reduces the endoperoxide to the corresponding alcohol (PGH2), the precursor of PGs, thromboxanes, and prostacyclins.1,2 It is now well established that there are two distinct isoforms of COX, COX-1 and COX-2. Cayman’s COX Fluorescent Inhibitor Screening Assay provides a convenient fluorescence-based method for screening ovine COX-1 and human recombinant COX-2 for isozyme-specific inhibitors. The assay utilizes the peroxidase component of COXs. In this assay, the reaction between PGG2 and ADHP (10-acetyl-3,7-dihydroxyphenoxazine) produces the highly fluorescent compound resorufin. Resorufin fluorescence can be analyzed with an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm.
1
Nugteren, D.H., and Hazelhof, E. Isolation and properties of intermediates in prostaglandin biosynthesis. Biochim Biophys Acta 326 448-461 (1973).
2
Hamberg, M., and Samuelsson, B. Detection and isolation of an endoperoxide intermediate in prostaglandin biosynthesis. Proc Natl Acad Sci USA 70 899-903 (1973).
Xie, W., Chipman, J.G., Robertson, D.L., et al. Expression of a mitogen-responsive gene encoding prostaglandin synthase is regulated by mRNA splicing. Proc Natl Acad Sci USA 88 2692-2696 (1991).
Kargman, S., Wong, E., Greig, G.M., et al. Mechanism of selective inhibition of human prostaglandin G/H synthase-1 and -2 in intact cells. Biochem Pharmacol 52 1113-1125 (1996).
Smith, C.J., Zhang, Y., Koboldt, C.M., et al. Pharmacological analysis of cyclooxygenase-1 in inflammation. Proc Natl Acad Sci USA 95 13313-13318 (1998).
Nugteren, D.H., and Hazelhof, E. Isolation and properties of intermediates in prostaglandin biosynthesis. Biochim Biophys Acta 326 448-461 (1973).
Hamberg, M., and Samuelsson, B. Detection and isolation of an endoperoxide intermediate in prostaglandin biosynthesis. Proc Natl Acad Sci USA 70 899-903 (1973).
Blobaum, A.L., and Marnett, L.J. Structural and functional basis of cyclooxygenase inhibition. J Med Chem 50(7) 1425-1441 (2007).
Jang, M., Cai, L., Udeani, G.O., et al. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275 218-220 (1997).
Show all 7
Hide all but first 3