This proposal is aimed at developing a reliable approach for basic mechanistic studies on the genotoxic effect of biological aldehyde at the molecular level. The primary effort is focused on unraveling the structural specificity, dynamics and quantitation of the interactions of pyruvaldehyde with DNA in vitro. The sites of reaction and the product distribution will be initially studied by a direct approach using solution and solid-state carbon-13 nuclear magnetic resonance spectroscopy, and then by an indirect approach using enzymic degradation followed by high performance liquid chromatographic separation and quantitation. The success of this proposal will pave the way for the in vivo studies on the formation and repair of potential mutagenic/carcinogenic DNA adducts in cells in culture and animals. The bioanalytical methodologies developed in this proposal can also be applied to the study of DNA modifications by other dicarbonyl compounds. It will provide the molecular basis for understanding the basic mutagenic/carcinogenic mechanisms of dicarbonyl compounds.
