A thermodynamic and spectroscopic study on the binding of berenil to poly d(AT) and to poly (dA) x poly (dT)

The complete thermodynamic profile for the non-intercalative binding of berenil to the alternating copolymer poly d(AT) and to the homopolymer poly (dA) x poly (dT) was investigated. Differential Scanning Calorimetry (DSC) and UV absorbance spectroscopy have been used to characterize and to compare the binding of berenil to the different synthetic polymers. Both double stranded DNA's show two types of binding; one stronger binding mode at low berenil concentrations and a weaker, in the case of poly d(AT)-berenil complexes slightly cooperative binding mode at higher drug to base pair ratios. For the interaction of berenil with poly d(AT) the thermodynamic data delta G(bind)0 = -33 kJ/mol drug, delta H(bind)0 = -29 kJ/mol of drug and delta S(bind)0 = +13 J/Kmol of drug were calculated. For the minor groove binding of berenil to poly (dA) x poly (dT) the following values were obtained: delta G(bind)0 = -34 kJ/mol of drug, delta H(bind)0 = -25 kJ/mol of drug and delta S(bind)0 = +30 J/Kmol of drug. Temperature-dependent UV absorbance spectroscopy revealed for both duplexes a biphasic "melting" behavior. However, the saturated nucleic acids (drug to base pair ratio 0.33) "melted" monophasically and with a decreased length of the cooperative unit. The obtained apparent equilibrium constants K(app) for the complexation with the discharged drug molecule showed to be a sensitive function of the ionic environment. But in contradiction to the expected release of two counterions into the solvent only a value of 1.0 was observed for the alternating copolymer poly d(AT). The complexation of berenil with poly (dA) x poly (dT) is followed by a release of 1.4 ions indicating stronger electrostatic interactions. For both polynucleotides the predicted release of two ions is not achieved. This is due to the presence of a binding mode, which involves less electrostatic interactions. From the complete data set it is proposed that the mode of binding is closely related to that found for the analogue minor groove binders DAPI and netropsin.