Identification and regulation of a new vertebrate cytochrome P450 subfamily, the CYP2Ps, and functional characterization of CYP2P3, a conserved arachidonic acid epoxygenase/19-hydroxylase.
Three genes cloned from Fundulus heteroclitus (killifish) define a new P450 subfamily, CYP2P. Structurally, the CYP2Ps are related to fish CYP2Ns and mammalian CYP2Js. CYP2P transcripts are expressed predominantly in liver and intestine. CYP2P3 coexpressed with P450 oxidoreductase in a baculovirus system catalyzed benzphetamine-N-demethylation and arachidonic acid oxidation, forming 14,15-, 11,12-, and 8,9-epoxyeicosatrienoic acids and 19-hydroxyeicosatetraenoic acid. CYP2P3 regio- and enantioselectivities with arachidonic acid were remarkably similar to human CYP2J2 and rat CYP2J3. Epoxyeicosatrienoic acids and their corresponding hydration products, the dihydroxyeicosatrienoic acids, were detected in killifish liver and intestine, indicating metabolism of arachidonic acid by killifish P450s in vivo. Levels of these products in killifish intestine were higher than those in mammalian intestine. 12-O-Tetradecanoyl phorbol 13-acetate suppressed expression of CYP2P2 and CYP2P3 in killifish intestine; fasting itself suppressed expression of CYP2P2/3 but not CYP2P1. In rat intestine fasting similarly depressed the levels of CYP2J proteins. The CYP2Ps and the CYP2Js appear to be derived from a common ancestral gene, likely a fatty acid monooxygenase.
Oleksiak
Marjorie F
MF
Biology Department, Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.
Wu
Shu
S
Parker
Carol
C
Qu
Wei
W
Cox
Rachel
R
Zeldin
Darryl C
DC
Stegeman
John J
JJ
D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001589:Benzphetamine / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D005215:Fasting; D023781:Fundulidae / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D009928:Organ Specificity; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D016415:Sequence Alignment; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014714:Vertebrates / Q000235:genetics*
C107016:14,15-dihydroxyeicosatrienoic acid; D006893:Hydroxyeicosatetraenoic Acids; D012333:RNA, Messenger; D001589:Benzphetamine; D016718:Arachidonic Acid; C046783:11,12-epoxy-5,8,14-eicosatrienoic acid; C046782:14,15-epoxy-5,8,11-eicosatrienoic acid; D003577:Cytochrome P-450 Enzyme System; C473194:CYP2P3 protein, Fundulus heteroclitus; D015126:8,11,14-Eicosatrienoic Acid; D013755:Tetradecanoylphorbol Acetate