Philip Kingsley
Last active: 3/12/2020

Characterization of an AM404 analogue, N-(3-hydroxyphenyl)arachidonoylamide, as a substrate and inactivator of prostaglandin endoperoxide synthase.

Turman MV, Kingsley PJ, Marnett LJ
Biochemistry. 2009 48 (51): 12233-41

PMID: 19928795 · PMCID: PMC2797370 · DOI:10.1021/bi901181z

N-(4-Hydroxyphenyl)arachidonoylamide (AM404) is an inhibitor of endocannabinoid inactivation that has been used in cellular and animal studies. AM404 is a derivative of arachidonic acid and has been reported to inhibit arachidonate oxygenation by prostaglandin endoperoxide synthase-1 and -2 (PGHS-1 and -2, respectively). While examining the structural requirements for inhibition of PGHS, we discovered that the meta isomer of AM404, N-(3-hydroxyphenyl)arachidonoylamide (3-HPAA), is a substrate for purified PGHS. PGHS-2 efficiently oxygenated 3-HPAA to prostaglandin and hydroxyeicosatetraenoate products. No oxidation of the phenolamide moiety was observed. 3-HPAA appeared to be converted by PGHS-1 in a similar manner; however, conversion was less efficient than that by PGHS-2. PGHS-2 was selectively, dose-dependently, and irreversibly inactivated in the presence of 3-HPAA. Complete inactivation of PGHS-2 was achieved with 10 muM 3-HPAA. Preliminary characterization revealed that 3-HPAA inactivation did not result from covalent modification of PGHS-2 or damage to the heme moiety. These studies provide additional insight into the structural requirements for substrate metabolism and inactivation of PGHS and report the first metabolism-dependent, selective inactivator of PGHS-2.

MeSH Terms (9)

Animals Arachidonic Acids Cyclooxygenase 1 Cyclooxygenase 2 Cyclooxygenase Inhibitors Kinetics Mice Sheep Substrate Specificity

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