PMID: 8950354Nov 4, 1996Paper

Monophosphate 32P-postlabeling assay of DNA adducts from 1,2:3,4-diepoxybutane, the most genotoxic metabolite of 1,3-butadiene: in vitro methodological studies and in vivo dosimetry

Mutation Research
N MabonK Randerath

Abstract

Among the main DNA-reactive metabolites of 1,3-butadiene (BD), both 1,2:3,4-butadiene diepoxide (BDE) and 1,2-epoxy-3-butene (BME) have been reported in mice and rats exposed to BD, but blood and tissue levels of these metabolites are much higher in mice than in rats under similar exposure conditions. BDE, being more reactive and genotoxic than BME, is thought to be responsible for the greater susceptibility of mice to BD carcinogenicity. While BDE is a DNA-alkylating agent and some BDE adducts have been characterized, no sufficiently sensitive method has been reported for studying BDE-DNA binding in vivo. In the present investigation, a modified dinucleotide/monophosphate version of the 32P-postlabeling assay was applied to detect BDE-DNA adducts, which were prepared by reacting BDE with calf thymus DNA or deoxyribooligonucleotides [(AC)10, (AG)10, (CCT)7 and (GGT)7] in vitro or with skin DNA of mice in vivo upon topical treatment. Optimal resolution by 2-D PEI-cellulose TLC of the highly polar 5'-monophosphate adducts was achieved at +4 degrees C using 0.3 M LiCI (DI) and 0.4 M NaCl, 0.04 M H3BO3, pH 7.6 (D2). The profiles of the 32P-postlabeled adducts were similar for calf thymus and skin DNA, with 3 major spots being detec...Continue Reading

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Citations

Jun 9, 2001·Chemico-biological Interactions·K HughesM Walker
Jul 27, 2001·Mutation Research·G D ZhouK Randerath
Feb 18, 2003·Journal of Toxicology and Environmental Health. Part B, Critical Reviews·K HughesR Beauchamp
Sep 28, 2010·Critical Reviews in Toxicology·Richard J AlbertiniMichael L Gargas
Dec 15, 2015·Mutation Research. Genetic Toxicology and Environmental Mutagenesis·Hueiwang Anna JengWen-Yi Lin
Sep 24, 2004·International Journal of Cancer. Journal International Du Cancer·Guo-Dong ZhouAnil K Jaiswal
Oct 20, 2007·Environmental and Molecular Mutagenesis·Guo-Dong ZhouKurt Randerath
Jul 20, 1999·Chemical Research in Toxicology·H KocJ A Swenberg

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