Comparison of rainbow trout and mammalian cytochrome P450 enzymes: evidence for structural similarity between trout P450 LMC5 and human P450IIIA4.
Studies were undertaken to determine the immunochemical relationship between constitutive trout cytochrome P450s and mammalian cytochrome P450IIIA enzymes. Polyclonal antibodies (IgG) generated against trout P450 LMC5 reacted strongly with P450IIIA1 in dexamethasone-induced rat liver microsomes and with P450IIIA4 in human liver microsomes in immunoblots. In contrast, rabbit anti-P450 LMC1 IgG did not recognize these proteins in rat and human liver microsomes. Reciprocal immunoblots using anti-rat P450IIIA1 showed that this antibody does not recognize trout P450 LMC1 or LMC5. However, anti-human P450IIIA4 IgG was found to cross react strongly with P450 LMC1 and LMC5. Progesterone 6 beta-hydroxylase activity of trout liver microsomes, a reaction catalyzed by P450 LMC5, was markedly inhibited by anti-P450IIIA4 and by gestodene, a mechanism-based inactivator of P450IIIA4. These results provide evidence for a close structural similarity between trout P450 LMC5 and human P450IIIA4.
Miranda
C L
CL
Department of Agricultural Chemistry, Oregon State University, Corvallis 97331.
Wang
J L
JL
Henderson
M C
MC
Zhao
X
X
Guengerich
F P
FP
Buhler
D R
DR
D000818:Animals; D003429:Cross Reactions; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000276:immunology; D003907:Dexamethasone / Q000494:pharmacology; D006900:Hydroxylation; D007074:Immunoglobulin G / Q000276:immunology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000032:analysis / Q000276:immunology; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000378:metabolism; D008970:Molecular Weight; D009652:Norpregnenes / Q000494:pharmacology; D011374:Progesterone / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013329:Structure-Activity Relationship; D014337:Trout
D007074:Immunoglobulin G; D007527:Isoenzymes; D009652:Norpregnenes; C033273:Gestodene; D011374:Progesterone; D003907:Dexamethasone; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases