Energetics of the binding of phototoxic and cytotoxic plant alkaloid sanguinarine to DNA: Isothermal titration calorimetric studies

Abstract

Sanguinarine is a medically important plant alkaloid with remarkable pharmacological and biological activities. DNA binding is considered to be an important aspect in its mode of action. Isothermal titration calorimetric technique was used for the first time to derive the thermodynamic aspects of the interaction of the phototoxic and cytotoxic plant alkaloid sanguinarine with natural calf thymus DNA and four synthetic DNAs of differing base pair sequences under various environmental conditions The affinity of binding of sanguinarine (iminium form) was found to be in the order of 10 5 M −1 to calf thymus DNA. The binding affinity to calf thymus DNA decreased with increase of ionic strength and temperature. The salt dependence of the binding data showed that release of 0.55 U of the cations per bound alkaloid. The binding of the iminium form was exothermic under all conditions, but the uncharged alkanolamine form showed no binding to DNA. The heat capacity changes obtained from the temperature dependence of enthalpy indicated a value of −140 cal/mol K. Sanguinarine showed high specificity to alternating purine–pyrimidine sequences with affinity of the order 10 6 M −1 and the affinity to the polynucleotides varied in the order poly(dG-dC)·poly(dG-dC) > poly(dA-dT)·poly(dA-dT) > poly(dA)·poly(dT) > poly(dG)·poly(dC). The binding to alternating GC polymer was exothermic and enthalpy driven, to the homo GC polymer was exothermic and favoured by both negative enthalpy and positive entropy changes, to the alternating AT polymer was exothermic and enthalpy driven while to the homo AT polymer was endothermic and entropy driven.