Identification and Quantitation of Benzo[a]pyrene-DNA Adducts Formed in Mouse Skin

Eleanor G Rogan, P. D. Devanesan, N. V S RamaKrishna, S. Higginbotham, N. S. Padmavathi, K. Chapman, Ercole Cavalieri, H. Jeong, R. Jankowiak, G. J. Small

Research output: Contribution to journalArticle

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Abstract

The DNA adducts of benzo[a]pyrene (BP) formed in vitro were previously identified and quantitated. In this paper, we report the identification and quantitation of the depurination adducts of BP, 8-(benzo[a]pyren-6-yl)guanine (BP-6-C8Gua), BP-6-N7Gua, and BP-6-N7Ade, formed in mouse skin by one-electron oxidation, as well as the major stable adduct formed via the diolepoxide pathway, BP diolepoxide bound at C-10 to the 2-amino of dG (BPDE-10-N2dG). Identification of the depurination adducts was achieved by HPLC and fluorescence line narrowing spectroscopy. The depurination adducts, BP-6-C8Gua (34%), BP-6-N7Gua (10%), and BP-6-N7Ade (30%), constituted 74% of the adducts found in mouse skin 4 h after treatment with BP. The stable adduct BPDE-10-N2dG accounted for 22% of the adducts. Treatment of the skin with BP-7,8-dihydrodiol or BP diolepoxide yielded almost exclusively the stable adduct BPDE-10-N2dG. When BP or BP-7,8-dihydrodiol was bound to RNA or denatured DNA in reactions catalyzed by rat liver microsomes, no depurination adducts were detected. The profiles of stable adducts were similar both qualitatively and quantitatively with native or denatured DNA. With activation of BP by horseradish peroxidase, the profiles of stable adducts differed with native and denatured DNA. The total amount of adducts with denatured DNA was only 25% of the amount detected with native DNA. No depurination adducts were detected with denatured DNA or RNA in the peroxidase system. The results reported here demonstrate that in mouse skin BP-DNA adducts are predominantly formed by one-electron oxidation and that this mechanism of activation requires double helical DNA for formation of adducts.

Original languageEnglish (US)
Pages (from-to)356-363
Number of pages8
JournalChemical Research in Toxicology
Volume6
Issue number3
DOIs
StatePublished - Jan 1 1993

Fingerprint

Skin
7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide
DNA
DNA Adducts
RNA
Electrons
Chemical activation
Guanine
Oxidation
Liver Microsomes
Horseradish Peroxidase
Peroxidase
benzo(a)pyrene-DNA adduct
Spectrum Analysis
Liver
Fluorescence
Rats
High Pressure Liquid Chromatography
Spectroscopy
7-(benzo(a)pyren-6-yl)guanine

ASJC Scopus subject areas

  • Toxicology

Cite this

Rogan, E. G., Devanesan, P. D., RamaKrishna, N. V. S., Higginbotham, S., Padmavathi, N. S., Chapman, K., ... Small, G. J. (1993). Identification and Quantitation of Benzo[a]pyrene-DNA Adducts Formed in Mouse Skin. Chemical Research in Toxicology, 6(3), 356-363. https://doi.org/10.1021/tx00033a017

Identification and Quantitation of Benzo[a]pyrene-DNA Adducts Formed in Mouse Skin. / Rogan, Eleanor G; Devanesan, P. D.; RamaKrishna, N. V S; Higginbotham, S.; Padmavathi, N. S.; Chapman, K.; Cavalieri, Ercole; Jeong, H.; Jankowiak, R.; Small, G. J.

In: Chemical Research in Toxicology, Vol. 6, No. 3, 01.01.1993, p. 356-363.

Research output: Contribution to journalArticle

Rogan, EG, Devanesan, PD, RamaKrishna, NVS, Higginbotham, S, Padmavathi, NS, Chapman, K, Cavalieri, E, Jeong, H, Jankowiak, R & Small, GJ 1993, 'Identification and Quantitation of Benzo[a]pyrene-DNA Adducts Formed in Mouse Skin', Chemical Research in Toxicology, vol. 6, no. 3, pp. 356-363. https://doi.org/10.1021/tx00033a017
Rogan EG, Devanesan PD, RamaKrishna NVS, Higginbotham S, Padmavathi NS, Chapman K et al. Identification and Quantitation of Benzo[a]pyrene-DNA Adducts Formed in Mouse Skin. Chemical Research in Toxicology. 1993 Jan 1;6(3):356-363. https://doi.org/10.1021/tx00033a017
Rogan, Eleanor G ; Devanesan, P. D. ; RamaKrishna, N. V S ; Higginbotham, S. ; Padmavathi, N. S. ; Chapman, K. ; Cavalieri, Ercole ; Jeong, H. ; Jankowiak, R. ; Small, G. J. / Identification and Quantitation of Benzo[a]pyrene-DNA Adducts Formed in Mouse Skin. In: Chemical Research in Toxicology. 1993 ; Vol. 6, No. 3. pp. 356-363.
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abstract = "The DNA adducts of benzo[a]pyrene (BP) formed in vitro were previously identified and quantitated. In this paper, we report the identification and quantitation of the depurination adducts of BP, 8-(benzo[a]pyren-6-yl)guanine (BP-6-C8Gua), BP-6-N7Gua, and BP-6-N7Ade, formed in mouse skin by one-electron oxidation, as well as the major stable adduct formed via the diolepoxide pathway, BP diolepoxide bound at C-10 to the 2-amino of dG (BPDE-10-N2dG). Identification of the depurination adducts was achieved by HPLC and fluorescence line narrowing spectroscopy. The depurination adducts, BP-6-C8Gua (34{\%}), BP-6-N7Gua (10{\%}), and BP-6-N7Ade (30{\%}), constituted 74{\%} of the adducts found in mouse skin 4 h after treatment with BP. The stable adduct BPDE-10-N2dG accounted for 22{\%} of the adducts. Treatment of the skin with BP-7,8-dihydrodiol or BP diolepoxide yielded almost exclusively the stable adduct BPDE-10-N2dG. When BP or BP-7,8-dihydrodiol was bound to RNA or denatured DNA in reactions catalyzed by rat liver microsomes, no depurination adducts were detected. The profiles of stable adducts were similar both qualitatively and quantitatively with native or denatured DNA. With activation of BP by horseradish peroxidase, the profiles of stable adducts differed with native and denatured DNA. The total amount of adducts with denatured DNA was only 25{\%} of the amount detected with native DNA. No depurination adducts were detected with denatured DNA or RNA in the peroxidase system. The results reported here demonstrate that in mouse skin BP-DNA adducts are predominantly formed by one-electron oxidation and that this mechanism of activation requires double helical DNA for formation of adducts.",
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AU - Rogan, Eleanor G

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AU - Higginbotham, S.

AU - Padmavathi, N. S.

AU - Chapman, K.

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N2 - The DNA adducts of benzo[a]pyrene (BP) formed in vitro were previously identified and quantitated. In this paper, we report the identification and quantitation of the depurination adducts of BP, 8-(benzo[a]pyren-6-yl)guanine (BP-6-C8Gua), BP-6-N7Gua, and BP-6-N7Ade, formed in mouse skin by one-electron oxidation, as well as the major stable adduct formed via the diolepoxide pathway, BP diolepoxide bound at C-10 to the 2-amino of dG (BPDE-10-N2dG). Identification of the depurination adducts was achieved by HPLC and fluorescence line narrowing spectroscopy. The depurination adducts, BP-6-C8Gua (34%), BP-6-N7Gua (10%), and BP-6-N7Ade (30%), constituted 74% of the adducts found in mouse skin 4 h after treatment with BP. The stable adduct BPDE-10-N2dG accounted for 22% of the adducts. Treatment of the skin with BP-7,8-dihydrodiol or BP diolepoxide yielded almost exclusively the stable adduct BPDE-10-N2dG. When BP or BP-7,8-dihydrodiol was bound to RNA or denatured DNA in reactions catalyzed by rat liver microsomes, no depurination adducts were detected. The profiles of stable adducts were similar both qualitatively and quantitatively with native or denatured DNA. With activation of BP by horseradish peroxidase, the profiles of stable adducts differed with native and denatured DNA. The total amount of adducts with denatured DNA was only 25% of the amount detected with native DNA. No depurination adducts were detected with denatured DNA or RNA in the peroxidase system. The results reported here demonstrate that in mouse skin BP-DNA adducts are predominantly formed by one-electron oxidation and that this mechanism of activation requires double helical DNA for formation of adducts.

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