Heme prosthetic group required for acetylation of prostaglandin H synthase by aspirin.
The ability of aspirin to acetylate PGH synthase was determined by reacting [3H-acetyl]-aspirin with purified enzyme followed by high pressure liquid chromatography analysis of the protein components of the reaction mixture. Heme-reconstituted enzyme incorporated approximately one acetyl group per 70-kDa subunit, whereas apoprotein incorporated 0.1 acetyl group per subunit. The ability of the heme prosthetic group to enhance acetylation of the protein was correlated with its ability to protect the Arg253-Gly254 peptide bond from cleavage by trypsin. Thus, heme-induced alteration of protein conformation may contribute to the enhanced labeling of Ser506 by aspirin. The present results indicate that irreversible inactivation of prostaglandin H synthase by aspirin occurs only when the heme prosthetic group is bound to the protein. Considering its short in vivo half-life, it is likely that aspirin inactivates only the steady-state fraction of PGH synthase in a cell that is active but not newly synthesized apoprotein. This may contribute to the differential kinetics of inactivation and recovery of PGH synthase activity in platelets and vascular endothelial cells after administration of low dose aspirin as a prophylactic agent against cardiovascular disease.
Chen
Y N
YN
Department of Chemistry, Wayne State University, Detroit, Michigan 48202.
Marnett
L J
LJ
D000107:Acetylation; D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D001241:Aspirin / Q000502:physiology*; D016861:Cyclooxygenase Inhibitors; D006418:Heme / Q000502:physiology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013552:Swine; D014357:Trypsin / Q000494:pharmacology
D001059:Apoproteins; D016861:Cyclooxygenase Inhibitors; D006418:Heme; D011451:Prostaglandin-Endoperoxide Synthases; D014357:Trypsin; D001241:Aspirin