Interactions of a fragment of bleomycin with deoxyribodinucleotides: nuclear magnetic resonance studies.

Proton NMR spectroscopy was used to establish certain geometrical parameters of the complexes formed between N-(3-aminopropyl)-2'-(2-acetamidoethyl)-2,4'-bithiazole-4-carboxamide hydrochloride (BLMF), a fragment of bleomycin, and various deoxyribodinucleotides. All proton resonances in these compounds have been assigned; chemical shifts were recorded as functions of their concentration. In the complex formed between BLMF and pdG-dC, chemical shifts of the bithiazole protons (measured with respect to values extrapolated to infinite dilution) were displaced upfield by 0.4 ppm. Other proton resonances of BLMF were shifted upfield but to a lesser extent. After corrections are made for self-stacking, maximum values for induced chemical shifts of the bithiazole protons are reached at a dinucleotide/BLMF ratio of 2. Coupling sums for dinucleotides (12.5-13.7 Hz) were unchanged following complexation, suggesting that there is no marked change in sugar conformation when BLMF is bound. On the basis of these results and of molecular model building studies, we propose a three-dimensional structure for a BLMF:pdG-dc complex in which the thiazole rings are intercalated in the duplex and stack preferentially on the purines. Projected on the same plane, the horizontal axis connecting the center of both bithiazole rings in this configuration superimposes on the axis connecting the centers of the purine bases. In this complex, both thiazole protons extend into the minor groove and the positively charged terminal amine binds to the negatively charged phosphate group of DNA.