title: Metabolism of carcinogenic heterocyclic and aromatic amines by recombinant human cytochrome P450 enzymes.
abstract: The N-hydroxylation of carcinogenic arylamines represents an initial step in their metabolic activation. Animal studies have shown that this reaction is catalyzed by the cytochrome P450 (P450) enzymes P450 1A1 and P450 1A2. In this study, utilizing enzymes expressed in Escherichia coli (and purified) or in human B-lymphoblastoid cells, the catalytic activities of recombinant human P450 1A1, P450 1A2, and P450 3A4 for N-hydroxylation of several carcinogenic arylamines were determined. P450 1A2 from both expression systems catalyzed the N-hydroxylation of 4-aminobiphenyl and the heterocyclic amines, 2-amino-3-methylimidazo[4,5-f/quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Rates were similar, with values of 1.1-7.8 nmol/min/nmol P450. In contrast, P450 1A1 catalyzed N-hydroxylation of only PhIP, and no activity was observed with P450 3A4. Further kinetic analysis with purified P450 1A2 showed similar Km and Vmax values for N-hydroxylation of the arylamines. Furafylline and fluvoxamine, inhibitors of P450 1A2 activity in human liver microsomes, were found to be inhibitory of the recombinant P450 1A2 N-hydroxylation activity. Results from this study are supportive of a major role for human P450 1A2 in the metabolic activation of arylamines.
authors.affiliation: National Center for Toxicology Research, Jefferson, AR 72079, USA.
authors.forename: G J
authors.initials: GJ
authors.lastname: Hammons
authors.forename: D
authors.initials: D
authors.lastname: Milton
authors.forename: K
authors.initials: K
authors.lastname: Stepps
authors.forename: F P
authors.initials: FP
authors.lastname: Guengerich
authors.forename: R H
authors.initials: RH
authors.lastname: Tukey
authors.forename: F F
authors.initials: FF
authors.lastname: Kadlubar
mesh_terms: D000588:Amines / Q000378:metabolism*; D002273:Carcinogens / Q000378:metabolism*; D002384:Catalysis; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000588:Amines; D002273:Carcinogens; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System