32P-Postlabeling in Studies of PAH Activation.

摘要

Polycyclic aromatic hydrocarbons (PAH) undergo metabolic activation reactions to form intermediates that react with DNA to form covalent adducts. PAH administration leads to the formation of various types of DNA adducts that may differ between species, strains, and tissues due to differences in metabolic activation and repair. The identification of the structures of PAH-DNA adducts may be accomplished by three approaches: co-chromatography with synthetic mononucleotide adduct standards; examining the adducts resulting from metabolism of pathway intermediates; or by chemically blocking metabolic activation at specific sites on the PAH. Administration of putative metabolic intermediates of a PAH leads to enhanced formation of DNA adducts resulting from further activation along that pathway. Conversely, chemically blocking a bond or position on a PAH prevents adducts arising from activation at the site. By comparing the DNA adduct spectra generated by metabolite, blocked forms, and parent PAH administration, the pathways important in the metabolic activation of the PAH in each tissue may be deduced. Partial identification of these adducts may also be determined by co-chromatography with the reaction products of synthetic epoxides and defined polydeoxynucleotides, and more thorough identification by using synthetic DNA adduct standards. These approaches have all been successfully applied to studies of PAH activation, and are reviewed in the paper.