Purification and Characterization of Cytochrome P-450 Induced by Benz(a)anthracene in Mouse Skin Microsomes

Takeshi Ichikawa, Shin Ichi Hayashi, Mitsuhide Noshiro, Kazuaki Takada, Kyuichiro Okuda

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

Topical application of benz(a)anthracene to mouse skin elicited a 2-fold increase in cytochrome P-450 content, with accompanying increases in monooxygenase activities such as benzo(a)pyrene hydroxylation, 7-ethoxycoumarin 0-deethylation, and acetanilide 4-hydroxylation, in the microsomes. A major form of cytochrome P-450 was purified from skin microsomes of mice treated with polycyclic aromatic hydrocarbon. A specific content of 1.95 nmol/mg of protein, which corresponded to 48-fold purification from the ndcrosomes was observed. The purified protein produced a single major band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis having a molecular weight of 55,000. Using Western blotting, the band immunochemiadly cross-reacted with antibody which had been raised against rat liver cytochrome p-450mc-i.The purified preparation efficiently catalyzed benzo(a)pyrene hydroxylation and 7-ethoxycoumarin 0-deethylation when reconstituted with NADPH-cyto-chrome P-450 reductase. These activities were inhibited by 7,8-benzoflavone as well as anti-cytochrome p-450mc-iantibody, but not by p-450pb-iantibody. The results indicate that, in mouse skin microsomes, a cytochrome P-450 induced by benz(a)anthracene is enzymatically and immunochemically similar to rat liver cytochrome p-450mc-i-It is suggested that this enzyme plays an important role in the activation of carcinogenic polycyclic aromatic hydrocarbons.

Original languageEnglish
Pages (from-to)806-809
Number of pages4
JournalCancer Research
Volume49
Issue number4
Publication statusPublished - 1989 Feb 15
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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