Abstract
CYP2C enzymes epoxidize arachidonic acid (AA) to metabolites involved in the regulation of vascular and renal function. We tested the hypothesis that eicosapentaenoic acid (EPA), a n-3 polyunsaturated fatty acid, may serve as an alternative substrate. Human CYP2C8 and CYP2C9, as well as rat CYP2C11 and CYP2C23, were co-expressed with NADPH-CYP reductase in a baculovirus/insect cell system. The recombinant enzymes showed high EPA and AA epoxygenase activities and the catalytic efficiencies were almost equal comparing the two substrates. The 17,18-double bond was the preferred site of EPA epoxidation by CYPs 2C8, 2C11, and 2C23. 17(R),18(S)-Epoxyeicosatetraenoic acid was produced with an optical purity of about 70% by CYPs 2C9, 2C11, and 2C23 whereas CYP2C8 showed the opposite enantioselectivity. These results demonstrate that EPA is an efficient substrate of CYP2C enzymes and suggest that n-3 PUFA-rich diets may shift the CYP2C-dependent generation of physiologically active eicosanoids from AA- to EPA-derived metabolites.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Aryl Hydrocarbon Hydroxylases / metabolism
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Catalysis
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Cell Line
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Chromatography, High Pressure Liquid
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Cytochrome P-450 CYP2C8
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Cytochrome P-450 CYP2C9
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Cytochrome P-450 CYP2J2
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Cytochrome P-450 Enzyme System / classification*
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Cytochrome P-450 Enzyme System / genetics
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Cytochrome P-450 Enzyme System / metabolism*
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Cytochrome P450 Family 2
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Eicosapentaenoic Acid / chemistry
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Eicosapentaenoic Acid / metabolism*
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Humans
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Kinetics
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Protein Isoforms / classification
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Protein Isoforms / genetics
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Protein Isoforms / metabolism
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Rats
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Spodoptera
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Stereoisomerism
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Steroid 16-alpha-Hydroxylase / metabolism
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Substrate Specificity
Substances
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Cyp2c23 protein, rat
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Protein Isoforms
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cytochrome P-450 CYP2C subfamily
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Cytochrome P-450 Enzyme System
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Eicosapentaenoic Acid
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CYP2C9 protein, human
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Cytochrome P-450 CYP2C9
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Aryl Hydrocarbon Hydroxylases
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CYP2C11 protein, rat
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CYP2C8 protein, human
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Cytochrome P-450 CYP2C8
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Cytochrome P-450 CYP2J2
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Cytochrome P450 Family 2
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Steroid 16-alpha-Hydroxylase