Equine cytochrome P450 2C92: cDNA cloning, expression and initial characterization.
Substantial gaps exist in our knowledge of the metabolic clearance of therapeutic agents in horses. Accordingly, a cytochrome P450 monooxygenase in the 2C family was cloned from an equine liver, sequenced and expressed in a baculovirus expression system. Catalytic activities of the recombinant protein were measured with a number of substrates. The protein, assigned CYP2C92, displayed optimal catalytic activity with diclofenac using molar ratios of CYP2C92 to NADPH CYP450 reductase of 1:18. Addition of cytochrome b(5) to diclofenac incubations had no significant effect on metabolic turnover. CYP2C92 catalyzed diclofenac metabolism was 20-fold slower than the human counterpart, CYP2C9. CYP2C92 demonstrated comparable tolbutamide and (S)-warfarin hydroxylase activity compared to CYP2C9, upon addition of b(5) to the reactions. The results of this study demonstrate substantial interspecies differences in metabolism of substrates by CYP2C orthologues in the horse and human and support the need to fully characterize this enzyme system in equids.
DiMaio Knych
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K.L. Maddy Equine Analytical Chemistry Laboratory, California Animal Health and Food Safety Laboratory, School of Veterinary Medicine, University of California, West Health Science Drive, Davis, CA 95616, USA. hkknych@ucdavis.edu
DeStefano Shields
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C
Buckpitt
A R
AR
Stanley
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D000818:Animals; D055162:Biocatalysis; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics*; D004008:Diclofenac / Q000378:metabolism; D015870:Gene Expression; D006736:Horses / Q000033:anatomy & histology / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D013045:Species Specificity; D018411:Spodoptera / Q000166:cytology / Q000235:genetics; D013379:Substrate Specificity
D018076:DNA, Complementary; D004008:Diclofenac; D003577:Cytochrome P-450 Enzyme System