Mutation of serine-516 in human prostaglandin G/H synthase-2 to methionine or aspirin acetylation of this residue stimulates 15- R-HETE synthesis

• Published in: FEBS letters Vol. 342; no. 1; pp. 33 - 37
• Main Authors: Mancini, Joseph A., O'Neill, Gary P., Bayly, Christopher, Vickers, Philip J.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 28.03.1994; Elsevier
• Subjects: 11-HETE; 11-hydroxyeicosatetraenoic acid; 15-HETE; 15-hydroxyeicosatetraenoic acid; Acetylation; acetylsalicylic acid; Analytical, structural and metabolic biochemistry; Animals; ASA; Aspirin - metabolism; Aspirin - pharmacology; Biological and medical sciences; Cell Line; COX-2; Cyclooxygenase-2; dimethyl sulphoxide; DMSO; EDTA; Enzymes and enzyme inhibitors; ethylenediamine tetraacetic acid; Fundamental and applied biological sciences. Psychology; Humans; Hydroxyeicosatetraenoic Acids - biosynthesis; Methionine - chemistry; Microsomes - metabolism; Mutagenesis; Mutagenesis, Site-Directed; non-steroidal antiinflammatory drugs; NSAIDS; Oxidoreductases; PGHS; PGHS-2; prostaglandin; prostaglandin G/H synthase; Prostaglandin G/H synthase-2; Prostaglandin-Endoperoxide Synthases - chemistry; Prostaglandin-Endoperoxide Synthases - genetics; Prostaglandin-Endoperoxide Synthases - metabolism; Prostaglandins - biosynthesis; SDS-PAGE; Serine - chemistry; sodium dodecyl sulphate polyacrylamide gel electrophoresis; Vaccinia virus; COX-2; DMSO; PGHS; PGHS-2; 15-HETE; EDTA; ASA; Vaccinia virus; 11-HETE; SDS-PAGE; Mutagenesis; NSAIDS; PG; Cyclooxygenase-2; Prostaglandin G/H synthase-2; Human; Prostaglandin-endoperoxide synthase; Chordopoxvirinae; Molecular form; Intramolecular oxidoreductases; Enzyme; Orthopoxvirus; Prostaglandin-I synthase; HETE; Virus; Isomerases; Structure activity relation; Poxviridae; Oxidoreductases; Acetylation; Mutation
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(94)80579-2

Prostaglandin G/H synthase (PGHS) is a key enzyme in cellular prostaglandin (PG) synthesis and is the target of non-steroidal anti-inflammatory agents. PGHS occurs in two isoforms, termed PGHS-1 and PGHS-2. These isoforms differ in several respects, including their enzymatic activity following acetylation by aspirin. While PG synthesis by both isoforms is inhibited by aspirin, 15- R-hydroxyeicosatetraenoic acid (15- R-HETE) synthesis by PGHS-2, but not PGHS-1, is stimulated by preincubation with aspirin. We have mutated the putative aspirin acetylation site of hPGHS-2, and expressed the mutants in COS-7 cells using recombinant vaccinia virus. Enzyme activity and inhibitor sensitivity studies provide evidence that Ser 516 is the aspirin acetylation site of human PGHS-2 and that substitution of a methionine residue at this position can mimic the effects of aspirin acetylation on enzyme activity.