Reconstitution of rabbit liver microsomal N-nitrosopyrrolidine α-hydroxylase activity

G. D. McCoy, Dennis Koop

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The in vitro α-hydroxylation of N-nitrosopyrrolidine (NPYR) by both isolated rabbit liver microsomes and purified cytochrome P-450 isozymes was investigated. Microsomes from untreated rabbits catalyzed the α-hydroxylation of NPYR at rates similar to those reported previously for rats, mice, and hamsters. The effect of established inducers of microsomal P-450 caused complex changes in apparent rates of α-hydroxylation of NPYR which made interpretation of responses to inducer pretreatment difficult and suggested the participation of multiple cytochrome P-450 isozymes in the metabolism of NPYR. Partial inhibition of α-hydroxylase activity by antibodies against rabbit isozymes 2, 3a, and 5 indicated the participation of at least these three isozymes in microsomal catalysis. Reconstitution studies using purified rabbit isozymes 2, 3a, 3b, 3c, 4, and 6 indicated that isozymes 2, 3a, 4, and 6 possessed significant α-hydroxylase activity with isozymes 3a and 6 exhibiting the highest activity when assayed at 20 mM NPYR. As NPYR concentrations were decreased, the rates of catalysis for the reconstituted systems were differentially decreased such that isozyme 3a exhibited the highest activity at low NPYR concentrations. These data indicate that isozyme 3a is the preferred catalyst for the α-hydroxylation of NPYR at low substrate concentrations and suggest that conditions such as chronic ethanol consumption which lead to the induction of isozyme 3a in rabbits or its orthologue in other species can account for enhanced rates of α-hydroxylation and metabolic activation of NPYR.

Original languageEnglish (US)
Pages (from-to)3987-3992
Number of pages6
JournalCancer Research
Volume48
Issue number14
StatePublished - 1988
Externally publishedYes

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N-Nitrosopyrrolidine
Mixed Function Oxygenases
Isoenzymes
Rabbits
Liver
Hydroxylation
Catalysis
Cytochrome P-450 Enzyme System
Liver Microsomes
Microsomes
Cricetinae

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Reconstitution of rabbit liver microsomal N-nitrosopyrrolidine α-hydroxylase activity. / McCoy, G. D.; Koop, Dennis.

In: Cancer Research, Vol. 48, No. 14, 1988, p. 3987-3992.

Research output: Contribution to journalArticle

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