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N-Nitrosodialkylamine dealkylation in reconstituted systems containing cytochrome P-450 purified from phenobarbitaland β-naphthoflavone-treated rats

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Abstract

Five cytochrome P-450 forms were purified from livers of rats pretreated with phenobarbital (PB) or β-naphthoflavone (BNF), and the oxidative dealkylation of N-nitrosodialkylamines by the reconstituted cytochrome P-450 systems was measured. PB-II (P450IIB1) showed very high N-nitrosomethybutylamine (NMBA) debutylase activity, high NMBA demethylase activity and high N-nitrosomethyl-benzylamine (NMBeA) debenzylase activity, suggesting that the increase following PB treatment in hepatic microsomal NMBA debutylation and NMBeA debenzylation was due to the induction of PB-II. BNF-H (P450IA2) showed very high NMBA debutylase and high NMBeA debenzylase activities, and BNF-L (P450IA1) showed NMBA debutylase and high NMBeA debenzylase activities. These results suggested that the increase by BNF pretreatment in hepatic microsomal NMBA debutylation was due mainly to the induction of BNF-H and in some part to that of BNF-L. PB-II also showed very high dealkylation activity of lipophilic N-nitrosodialkylamines with long alkyl moieties. On the other hand, BNF-H dealkylated N-nitrosodipropylamine (NDPA), N-nitrosomethylbutylamine (NMBA) and N-nitrosoethylbutylamine (NEBA) at higher rates than N-nitrosodibutylamine (NDBA). BNF-L dealkylated NEBA at higher rates than NMBeA and NDBA. These results reveal that substrate specificity of each cytochrome P-450 form in N-nitrosodialkylamine metabolism is different from each other and several forms of cytochrome P-450 support each N-nitrosamine dealkylase activity in mammalians.

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Abbreviations

NDMA:

N-nitrosodimethylamine

NMBA:

N-nitrosomethylbutylamine

NMBeA:

N-nitrosomethylbenzylamine

PB:

phenobarbital

BNF:

β-naphthoflavone

NMEA:

N-nitrosomethylethylamine

NDPA:

N-nitrosodipropylamine

NEBA:

N-nitrosoethylbutylamine

NDEA:

N-nitrosodiethylamine

NDBA:

N-nitrosodibutylamine

PMSF:

phenylmethylsulfonyl fluoride

NPD:

nitrogen phosphate detector

PAGE:

polyacrylamide gel electrophoresis

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Author notes

  1. T. Kawanishi

    Present address: Division of Pharmacology, Biological Safety Research Center, National Institute of Hygienic Sciences, 1-18-1 Kamiyoga, Setagaya-ku, 158, Tokyo, Japan

Authors and Affiliations

  1. Division of Pharmacology, Biological Safety Research Center, National Institute of Hygienic Sciences, 1-18-1 Kamiyoga, Setagaya-ku, 158, Tokyo, Japan

    Y. Ohno, A. Takanaka & Y. Omori

  2. Department of Pharmacology, School of Medicine, Keio University, Shinanomachi 35, Shinjuku, 160, Tokyo, Japan

    S. Kawano, Y. Yamazoe & R. Kato

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  1. T. Kawanishi
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  5. Y. Yamazoe
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  6. R. Kato
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  7. Y. Omori
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Kawanishi, T., Ohno, Y., Takanaka, A. et al. N-Nitrosodialkylamine dealkylation in reconstituted systems containing cytochrome P-450 purified from phenobarbitaland β-naphthoflavone-treated rats. Arch Toxicol 66, 137–142 (1992). https://doi.org/10.1007/BF02342508

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  • DOI: https://doi.org/10.1007/BF02342508

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