Identification of CYP2C9 as a human liver microsomal linoleic acid epoxygenase

Alison J. Draper, Bruce D. Hammock

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

Leukotoxin (9,10-epoxy-12-octadecanoate) and isoleukotoxin (12,13-epoxy- 9-octadecenoate) are monoepoxides of linoleic acid, synthesized by a cytochrome P450 monooxygenase and possibly by an oxidative burst of inflammatory cells. Recent experiments in this laboratory have indicated that the toxicity of leukotoxin and isoleukotoxin is not due to these epoxides, but to the 9,10- and 12,13-diol metabolites. Leukotoxin and isoleukotoxin are metabolized primarily by the soluble epoxide hydrolase to form leukotoxin diol. Investigations with recombinant cytochrome P450 enzymes have demonstrated that leukotoxin and isoleukotoxin can be formed by these enzymes. This study used a combination of experimental approaches to identify the major cytochrome P450 enzyme in human liver involved in linoleic acid epoxidation. The kinetic paramenters were determined; the K(m) of linoleic acid epoxidation by pooled human liver microsomes was 170 μM and the V(max) was 58 pmol/mg/min. Correlation analysis was performed using individual samples of human liver microsomes, and the best correlation of linoleic acid epoxidation activity was with tolbutamide hydroxylase activity, CYP2C9. Recombinant CYP2C9 was the most active in linoleic acid epoxygenation, and antibody and chemical inhibition also indicated the importance of CYP2C9. This enzyme, therefore, may serve as a therapeutic target in the treatment of inflammation in order to reduce the amount of circulating leukotoxin/isoleukotoxin and their related diols. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)199-205
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume376
Issue number1
DOIs
StatePublished - Apr 1 2000

Keywords

  • 9,10-epoxy-12-octadecenoate
  • CYP2C9
  • Cytochrome P450
  • Fatty acid oxidation
  • Leukotoxin

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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