Formation and reduction of aryl and heterocyclic nitroso compounds and significance in the flux of hydroxylamines.
Cytochrome p450 (p450) 1A2 and NADPH-P450 reductase (NPR) catalyzed the oxidation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), with consumption of NADPH. The oxidation rate of NADPH by p450 1A2/NPR increased with time in the presence of IQ until depletion of NADPH. This unusual autocatalytic pattern of NADPH oxidation could be rationalized by formation of a nitroso derivative (IQ-N=O) and the subsequent reduction of the hydroxylamine (IQ-NHOH) and IQ-N=O, which would consume more NADPH. The formation of IQ-NHOH and IQ-N=O from IQ was confirmed using HPLC/MS. Reduction of IQ-NHOH and IQ-N=O was NPR-dependent but did not require p450. Autocatalytic NADPH oxidation was also observed in the oxidation of other heterocyclic and arylamines. However, the N-hydroxyl and nitroso oxidation products of 2-aminofluorene and 4-aminobiphenyl were reduced nonenzymatically by NADPH, and NPR did not catalyze the reactions. We simulated the enzymatic kinetic model for possible pathways for IQ metabolism, which included the formation of IQ-N=O, using some kinetic parameters obtained from the experimental results. In the kinetic model, we could reproduce the similar curvature for NADPH oxidation and the formation of IQ-N=O, and the reduction of IQ-NHOH and IQ-N=O is required to explain the observed results for NADPH oxidation. Our results support a role for nitroso derivatives of HAAs in the unusual autocatalytic NADPH oxidation and may have relevance in terms of possible toxicities of the nitroso derivatives. Both IQ-NHOH and IQ-N=O were mutagenic in a bacterial tester system devoid of p450 and NPR; the mutagenicity of both was decreased by expression of NPR, consistent with the reduction of these compounds observed with purified NPR.
Kim
Donghak
D
Department of Biochemistry and Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Kadlubar
Fred F
FF
Teitel
Candee H
CH
Guengerich
F Peter
FP
D002273:Carcinogens / Q000737:chemistry* / Q000633:toxicity; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D013058:Mass Spectrometry; D008962:Models, Theoretical; D009152:Mutagenicity Tests; D009249:NADP / Q000737:chemistry*; D009251:NADPH-Ferrihemoprotein Reductase / Q000494:pharmacology; D009603:Nitroso Compounds / Q000737:chemistry* / Q000633:toxicity; D010084:Oxidation-Reduction; D011804:Quinolines / Q000737:chemistry* / Q000633:toxicity; D012486:Salmonella typhimurium / Q000235:genetics
D002273:Carcinogens; D009603:Nitroso Compounds; D011804:Quinolines; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; D009249:NADP; D009251:NADPH-Ferrihemoprotein Reductase