Biosynthesis of 20-hydroxyeicosatetraenoic acid (20-HETE) and 12 (S)-HETE by renal cortical microsomes of the cynomolgus monkey.
Renal cortical microsomes of the cynomolgus monkey, Macaca fascicularis, were incubated with [14C] arachidonic acid in the presence of NADPH. The two main metabolites were identified as 20-hydroxyeicosatetraenoic acid (20-HETE) and 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE) by capillary GC-MS and by chiral phase HPLC. 12(S)-HETE was formed in highly variable amounts by the very same preparation of microsomes on different occasions. The microsomal biosynthesis of 12(S)-HETE was increased several-fold by addition of Ca++, NADPH (or NADH) and ATP, and the biosynthesis was reproducible in the presence of these agents. The biosynthesis was unaffected by proadifen (SKF-525A), however, and was inhibited by nordihydroguaiaretic acid. Some preparations of renal medullary microsomes and cytosol also contained 12(S)-lipoxygenase activity, which was not stimulated by Ca2+, NADPH and ATP and thus resembled the well-known platelet enzyme. Significant formation of 12(R)-HETE could not be detected. In conclusion, 20-HETE was identified as the main cytochrome P-450-derived metabolite and 12(S)-HETE as the main lipoxygenase product in microsomes of monkey renal cortex.
Oliw
E H
EH
Department of Pharmacology, University of Uppsala, Sweden.
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D006893:Hydroxyeicosatetraenoic Acids / Q000096:biosynthesis*; D007672:Kidney Cortex / Q000378:metabolism*; D007679:Kidney Medulla / Q000378:metabolism; D008252:Macaca fascicularis; D008861:Microsomes / Q000378:metabolism*; D009249:NADP / Q000378:metabolism
D001095:Arachidonic Acids; D006893:Hydroxyeicosatetraenoic Acids; D016718:Arachidonic Acid; D009249:NADP; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D002118:Calcium