Interactions of benzyl viologen with surface-bound single- and double-stranded DNA

The interactions of benzyl viologen (BV) with single- and double-stranded calf-thymus DNA immobilized onto gold electrodes have been studied by electrochemical methods. Benzyl viologen interacts electrostatically with both double-stranded (ds) and single-stranded (ss) DNA, and the strength of the interactions is dependent on ionic strength (mu). The dicationic form (BV2+) binds to dsDNA 9 times more strongly than the singly reduced form, BV*+, in a pH 7.4 Tris-HCl buffer solution at mu = 8.4 mM. BV2+ binds to ssDNA 5 times more strongly than the BV*+ form. From measurements at mu = 8.4 mM, a binding constant (K2+) of 2.0 (+/-0.2) x 10(4) M(-1) and a binding site size (s) of 1 base pair were obtained, respectively, for dsDNA. For ssDNA, at the same ionic strength, the values obtained for K and s were 3.6 (+/-0.4) x10(4) M(-1) and 2 nucleotides, respectively. The amount of BV bound, whether to dsDNA or ssDNA, decreased with increasing ionic strength. Whereas the binding rate of BV to both dsDNA and ssDNA immobilized onto gold electrodes is relatively low, once immobilized, it dissociates rapidly away from the electrode surface. The electron-transfer rate constant for BV is moderately fast at both dsDNA- and ssDNA-modified gold electrodes. The application of benzyl viologen as an electroactive indicator capable of differentiating between surface-immobilized single- and double-stranded DNA in denaturation/regeneration cycles has been explored.