@kaigrass

Thermodynamics of DNA binding and condensation: Isothermal titration calorimetry and electrostatic mechanism

, , and . J. Mol. Bio., 296 (4): 1053--1063 (2000)

Abstract

The thermodynamics of binding of the trivalent cations cobalt hexammine and spermidine to plasmid DNA was studied by isothermal titration calorimetry. Two stages were observed in the course of titration, the first attributed to cation binding and the second to DNA condensation. A standard calorimetric data analysis was extended by applying an electrostatic binding model, which accounted for most of the observed data. Both the binding and condensation reactions were entropically driven (T Delta S similar to + 10 kcal/mol cation) and enthalpically opposed (Delta H similar to + 1 kcal/mol cation). As predicted from their relative sizes, the binding constants of the cations were indistinguishable, but cobalt hexammine had a much greater DNA condensing capacity because it is more compact than spermidine. The dependence of both the free energy of cobalt hexammine binding and the critical. cobalt hexammine concentration for DNA condensation on temperature and monovalent cation concentration followed the electrostatic model quite precisely. The heat capacity changes of both stages were positive, perhaps reflecting both the temperature dependence of the dielectric constant of water and the burial of polar surfaces. DNA condensation occurred when about 67 percent of the DNA phosphate charge was neutralized by cobalt hexammine and 87 percent by spermidine. During condensation, the remaining DNA charge was neutralized.

Links and resources

Tags