Formation of 19(S)-, 19(R)-, and 18(R)-hydroxyeicosatetraenoic acids by alcohol-inducible cytochrome P450 2E1

摘要: When reconstituted with cytochrome b5 and NADPH P450 oxidoreductase, 2E1 metabolized lauric, stearic, oleic, linoleic, linolenic, arachidonic acid to multiple metabolites. Two major metabolites, accounting for 78% of the total metabolism, were produced acid. The Vmax metabolite formation from was 5 nmol/min/nmol an apparent Km 62 microM. Gas chromatography-mass spectrometry analysis identified two metabolites as monohydroxylated eicosatetraenoic acids (HETEs). HETE 19-hydroxyeicosatetraenoic (19-HETE) comprised 46% produced. second omega-2 hydroxylated (18-HETE) 32% product formed. Chiral demonstrated that 19-HETE 70% 19(S)-HETE 30% 19(R)-HETE. In contrast, 18-HETE essentially 100% R isomer. Approximately 18% coeluted epoxyeicosatrienoic (EET) standards. EET 56.4% 14,15-EET 43.6% a mixture 11,12-EET 8,9-EET. 5,6-EET not detected. Anti-P450 IgG inhibited metabolism by renal hepatic microsomes prepared acetone-treated rabbits. With cortex microsomes, 67 25%, respectively, antibody. Liver microsomal 87% 70%. Thus, under conditions where is induced, enzyme could contribute significantly omega-1