Investigation of benzo[a]pyrene-globin adducts

The nature of the adducts formed between benzo[a]pyrene (BP) and globin were investigated in animals treated with [3H]BP. Modification levels on globin were determined by radioactivity measurements. Since BP tetraols can be released from benzo[a]pyrene diol epoxide modified protein and DNA by acid treatment, globin samples were treated with acid, released tetraols separated by HPLC and quantitated by scintillation counting. In addition, acid released material was measured in a competitive enzyme linked immunosorbent assay (ELISA) using antibodies which recognize BP tetraols. Both measurements indicate that only 2% of bound radioactivity could be released as free BP tetraols. These studies indicate that benzo[a]pyrene diol epoxide may not be the major metabolite of BP involved in globin binding.