title: Metabolic activation of heterocyclic amines and other procarcinogens in Salmonella typhimurium umu tester strains expressing human cytochrome P4501A1, 1A2, 1B1, 2C9, 2D6, 2E1, and 3A4 and human NADPH-P450 reductase and bacterial O-acetyltransferase.
abstract: We investigated roles of different forms of cytochrome P450 (P450 or CYP) in the metabolic activation of heterocyclic amines (HCAs) and other procarcinogens to genotoxic metabolite(s) in the newly developed umu tester strains Salmonella typhimurium (S. typhimurium) OY1002/1A1, OY1002/1A2, OY1002/1B1, OY1002/2C9, OY1002/2D6, OY1002/2E1 and OY1002/3A4, which express respective human P450 enzymes and NADPH-cytochrome P450 reductase (reductase) and bacterial O-acetyltransferase (O-AT). These strains were established by introducing two plasmids into S. typhimurium TA1535, one carrying both P450 and the reductase cDNA in a bicistronic construct under control of an IPTG-inducible double tac promoter and the other, pOA102, carrying O-AT and umuC"lacZ fusion genes. Expression levels of CYP were found to range between 35 to 550 nmol/l cell culture in the strains tested. O-AT activities in different strains ranged from 52 to 125 nmol isoniazid acetylated/min/mg protein. All HCAs tested, and 2-aminoanthracene and 2-aminofluorene exhibited high genotoxicity in the OY1002/1A2 strain, and genotoxicity of 2-amino-3-methylimidazo [4,5-f]quinoline was detected in both the OY1002/1A1 and OY1002/1A2 strains. 3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]-indole and 3-amino-1-methyl-5H-pyrido[4,3-b]-indole were activated in the OY1002/1A1, OY1002/1B1, OY1002/1A2, and OY1002/3A4 strains. Aflatoxin B(1) exhibited genotoxicity in the OY1002/1A2, OY1002/1A1, and OY1002/3A4 strains. beta-Naphthylamine and benzo[a]pyrene did not exhibit genotoxicity in any of the strains. These results suggest that CYP1A2 is the major cytochrome P450 enzyme involved in bioactivation of HCAs.
authors.affiliation: Osaka Prefectural Institute of Public Health, Higashinari-ku, 537-0025, Osaka, Japan. ysoda@iph.pref.osaka.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Oda
authors.forename: P
authors.initials: P
authors.lastname: Aryal
authors.forename: T
authors.initials: T
authors.lastname: Terashita
authors.forename: E M
authors.initials: EM
authors.lastname: Gillam
authors.forename: F P
authors.initials: FP
authors.lastname: Guengerich
authors.forename: T
authors.initials: T
authors.lastname: Shimada
mesh_terms: D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D000588:Amines / Q000633:toxicity; D001191:Arylamine N-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002273:Carcinogens / Q000633:toxicity*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005449:Fluorenes; D006571:Heterocyclic Compounds / Q000633:toxicity*; D006801:Humans; D007527:Isoenzymes; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity*; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D000588:Amines; D002273:Carcinogens; D005449:Fluorenes; D006571:Heterocyclic Compounds; D007527:Isoenzymes; D009153:Mutagens; C012177:2-aminofluorene; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; D000123:Acetyltransferases; C047057:isoniazid acetyltransferase; C019736:N-hydroxyarylamine O-acetyltransferase; D001191:Arylamine N-Acetyltransferase