Structure elucidation of a 6-methylbenzo[a]pyrene-DNA adduct formed by horseradish peroxidase in vitro and mouse skin in vivo

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Abstract

Activation of polycyclic aromatic hydrocarbons (PAH) by horseradish peroxidase (HRP) with H2O2 has been studied as a model system for one-electron oxidation. This peroxidase has been used to catalyze binding of 6-[14C]methylbenzo[a]pyrene (BP-6-CH3) to DNA, which was purified, hydrolyzed to deoxyribonucleosides and analyzed by high pressure liquid chromatography (HPLC). The predominant hydrocarbon-DNA adduct observed was identified as BP-6-CH3 bound at the 6-methyl group to the 2-amino group of dG, confirming that activation by HRP occurs by one-electron oxidation. When DNA from mouse skin treated in vivo with [14C]BP-6-CH3 was purified, hydrolyzed and analyzed by HPLC, a profile was observed which was qualitatively similar to that from the peroxidase system. In particular, the identified adduct with the hydrocarbon bound at the 6-methyl group to the 2-amino group of dG was obtained. These results demonstrate that one-electron oxidation is the mechanism of activation by HRP for aromatic hydrocarbons and indicate that the same mechanism may occur in mouse skin, a target tissue for hydrocarbon carcinogenesis.

Original languageEnglish (US)
Pages (from-to)111-122
Number of pages12
JournalChemico-Biological Interactions
Volume47
Issue number1
DOIs
StatePublished - Oct 15 1983

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DNA Adducts
Horseradish Peroxidase
Hydrocarbons
High pressure liquid chromatography
Skin
Chemical activation
Electrons
Oxidation
Peroxidase
High Pressure Liquid Chromatography
Deoxyribonucleosides
Aromatic Hydrocarbons
DNA
Polycyclic Aromatic Hydrocarbons
Carcinogenesis
Tissue
6-methylbenzo(a)pyrene
In Vitro Techniques

Keywords

  • Carcinogenic activation
  • DNA adduct structure
  • Horseradish peroxidase
  • Mouse skin
  • One-electron oxidation
  • Polycyclic aromatic hydrocarbons

ASJC Scopus subject areas

  • Toxicology

Cite this

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title = "Structure elucidation of a 6-methylbenzo[a]pyrene-DNA adduct formed by horseradish peroxidase in vitro and mouse skin in vivo",
abstract = "Activation of polycyclic aromatic hydrocarbons (PAH) by horseradish peroxidase (HRP) with H2O2 has been studied as a model system for one-electron oxidation. This peroxidase has been used to catalyze binding of 6-[14C]methylbenzo[a]pyrene (BP-6-CH3) to DNA, which was purified, hydrolyzed to deoxyribonucleosides and analyzed by high pressure liquid chromatography (HPLC). The predominant hydrocarbon-DNA adduct observed was identified as BP-6-CH3 bound at the 6-methyl group to the 2-amino group of dG, confirming that activation by HRP occurs by one-electron oxidation. When DNA from mouse skin treated in vivo with [14C]BP-6-CH3 was purified, hydrolyzed and analyzed by HPLC, a profile was observed which was qualitatively similar to that from the peroxidase system. In particular, the identified adduct with the hydrocarbon bound at the 6-methyl group to the 2-amino group of dG was obtained. These results demonstrate that one-electron oxidation is the mechanism of activation by HRP for aromatic hydrocarbons and indicate that the same mechanism may occur in mouse skin, a target tissue for hydrocarbon carcinogenesis.",
keywords = "Carcinogenic activation, DNA adduct structure, Horseradish peroxidase, Mouse skin, One-electron oxidation, Polycyclic aromatic hydrocarbons",
author = "Rogan, {Eleanor G} and Alaeddin Hakam and Ercole Cavalieri",
year = "1983",
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T1 - Structure elucidation of a 6-methylbenzo[a]pyrene-DNA adduct formed by horseradish peroxidase in vitro and mouse skin in vivo

AU - Rogan, Eleanor G

AU - Hakam, Alaeddin

AU - Cavalieri, Ercole

PY - 1983/10/15

Y1 - 1983/10/15

N2 - Activation of polycyclic aromatic hydrocarbons (PAH) by horseradish peroxidase (HRP) with H2O2 has been studied as a model system for one-electron oxidation. This peroxidase has been used to catalyze binding of 6-[14C]methylbenzo[a]pyrene (BP-6-CH3) to DNA, which was purified, hydrolyzed to deoxyribonucleosides and analyzed by high pressure liquid chromatography (HPLC). The predominant hydrocarbon-DNA adduct observed was identified as BP-6-CH3 bound at the 6-methyl group to the 2-amino group of dG, confirming that activation by HRP occurs by one-electron oxidation. When DNA from mouse skin treated in vivo with [14C]BP-6-CH3 was purified, hydrolyzed and analyzed by HPLC, a profile was observed which was qualitatively similar to that from the peroxidase system. In particular, the identified adduct with the hydrocarbon bound at the 6-methyl group to the 2-amino group of dG was obtained. These results demonstrate that one-electron oxidation is the mechanism of activation by HRP for aromatic hydrocarbons and indicate that the same mechanism may occur in mouse skin, a target tissue for hydrocarbon carcinogenesis.

AB - Activation of polycyclic aromatic hydrocarbons (PAH) by horseradish peroxidase (HRP) with H2O2 has been studied as a model system for one-electron oxidation. This peroxidase has been used to catalyze binding of 6-[14C]methylbenzo[a]pyrene (BP-6-CH3) to DNA, which was purified, hydrolyzed to deoxyribonucleosides and analyzed by high pressure liquid chromatography (HPLC). The predominant hydrocarbon-DNA adduct observed was identified as BP-6-CH3 bound at the 6-methyl group to the 2-amino group of dG, confirming that activation by HRP occurs by one-electron oxidation. When DNA from mouse skin treated in vivo with [14C]BP-6-CH3 was purified, hydrolyzed and analyzed by HPLC, a profile was observed which was qualitatively similar to that from the peroxidase system. In particular, the identified adduct with the hydrocarbon bound at the 6-methyl group to the 2-amino group of dG was obtained. These results demonstrate that one-electron oxidation is the mechanism of activation by HRP for aromatic hydrocarbons and indicate that the same mechanism may occur in mouse skin, a target tissue for hydrocarbon carcinogenesis.

KW - Carcinogenic activation

KW - DNA adduct structure

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KW - Mouse skin

KW - One-electron oxidation

KW - Polycyclic aromatic hydrocarbons

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