Understanding the Effect of Carbonate Ion on Cisplatin Binding to DNA

Abstract

The role of carbonate in the binding of cis-diamminedichloroplatinum(II) to DNA was investigated in order to understand the potential involvement of carbonato-cisplatin species in the mechanism of action of platinum anticancer agents. Cisplatin was allowed to react with both double- and single-stranded DNA in carbonate, phosphate, and HEPES buffers, and the products were analyzed by agarose gel electrophoresis and enzymatic digestion/mass spectrometry, respectively. The data from these experiments demonstrate (1) that carbonate, like other biological nucleophiles, forms relatively inert complexes with platinum that inactivate cisplatin and (2) that the major cisplatin−DNA adduct formed is a bifunctional cross-link. These results are in accord with previous studies of cisplatin−DNA binding and reveal that the presence of carbonate has no consequence on the nature of the resulting adducts.