Using UHF-dielectrometry to study protein structural transitions.

Abstract

UHF-dielectrometry method is based on the following facts: i) there is dispersion (i.e. dependence on frequency) of the dielectric permeability epsilon; ii) bound and free water have remarkable different epsilon, mobility and dispersion regions; iii) conformational changes in a macromolecule lead to redistribution of free and bound water and to change of the amount of free water molecules. Choosing the working frequency in the region of dispersion of free water molecules (9.2 GHz) we can detect conformational changes in proteins using free water as a marker. In this work the temperature dependencies of dielectric parameters of albumin and fibrinogen solutions were obtained in the temperature interval 5-40 degrees C. In contrast to dependencies for poor solvent, temperature dependencies of dielectric parameters for protein solutions are of non-monotonous character; they have a number of peculiarities in the temperature ranges of 8-10, 22-24 and 34-36 degrees C. At these temperatures redistribution of free and bound water in protein-water system occurs due to structural changes in protein molecules. In this work the mechanism of temperature changes of spatial organisation of protein molecules was proposed. Perhaps, this mechanism is responsible for maintenance of thermal stability of the functionally active conformation of native proteins.