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

FEBS Letters

Mancini, Joseph A.¹  Vickers, Philip J.¹  Bayly, Christopher²  O'Neill, Gary P.¹ 

[1]Department of Biochemistry and Molecular Biology, Merck Frosst Centre for Therapeutic Research, PO Box 1005, Pointe Claire-Dorval, Kirkland, Que., H9R 4P8, Canada

[2]Department of Biochemistry and Medicinal Chemistry, Merck Frosst Centre for Therapeutic Research, PO Box 1005, Pointe Claire-Dorval, Kirkland, Que., H9R 4P8, Canada

关键词: Prostaglandin G/H synthase-2;    PGHS-2;    Cyclooxygenase-2;    COX-2;    15-HETE;    Mutagenesis;    Vaccinia virus;    ASA;    acetylsalicylic acid;    DMSO;    dimethyl sulphoxide;    EDTA;    ethylenediamine tetraacetic acid;    11-HETE;    11-hydroxyeicosatetraenoic acid;    15-HETE;    15-hydroxyeicosatetraenoic acid;    NSAIDS;    non-steroidal antiinflammatory drugs;    PG;    prostaglandin;    PGHS;    prostaglandin G/H synthase;    SDS-PAGE;    sodium dodecyl sulphate polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(94)80579-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 Ser⁵¹⁶ 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.

【 授权许可】

Unknown