Metabolic activation of the pneumotoxin, 3-methylindole, by vaccinia-expressed cytochrome P450s.
Twelve human cytochrome P450s and one mouse P450 were produced in HepG2 cells using vaccinia virus cDNA expression and analyzed for their ability to bioactivate the pneumotoxin, 3-methylindole (3MI), to an electrophilic metabolite(s) which alkylated cellular macromolecules. Cell lysates containing CYP2C8, CYP3A4, CYP2A6 and CYP2F1 metabolized 3MI to an intermediate(s) that became covalently bound to lysate material. A control lysate produced from cells which had been infected with a wild-type vaccinia virus was not able to bioactivate 3MI. The mouse 1A2 enzyme metabolized 3MI at a rate of 75.4 pmol/mg protein/minute, while the rate of metabolism in the lysate containing the human 1A2 P450 enzyme was not different from that in the control lysate. Therefore, the catalytic capabilities of orthologous P450 enzymes to activate 3MI cannot be extrapolated among different species. These results indicate that human P450s are capable of bioactivating 3MI to a metabolite which binds to cellular macromolecules suggesting that this compound may be toxic to humans.
Thornton-Manning
J R
JR
Department of Pharmacology and Toxicology, University of Utah, Salt Lake City 84112.
Ruangyuttikarn
W
W
Gonzalez
F J
FJ
Yost
G S
GS
D000818:Animals; D001711:Biotransformation; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D011994:Recombinant Proteins / Q000378:metabolism; D012862:Skatole / Q000378:metabolism*; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics
D007527:Isoenzymes; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D012862:Skatole; D051544:Cytochrome P-450 CYP3A; C000592551:cytochrome P450 3A4, mouse