Spin labeling -- electron paramagnetic resonance spectroscopy study of temperature and salt dependent serum albumin stability

TL;DR

This study uses electron paramagnetic resonance to investigate how temperature, salt, and other factors affect the stability and conformational states of human serum albumin, revealing phase transitions and structural dynamics.

Contribution

It introduces a method to measure protein rotational dynamics and correlates it with stability and phase behavior under various conditions.

Findings

Correlation time relates to protein volume and stability.

Temperature and salt induce conformational phase transitions.

Phase diagrams indicate conditions for liquid-liquid transitions.

Abstract

Electron paramagnetic resonance signal parameters of TEMPO dichlorotriazine and TEMPO maleimide spin labels attached to protein human serum albumin molecules have been monitored at various temperatures, salts and sucrose concentrations. The method for measuring the correlation time of the protein rotational dynamics using viscosity isotherms was utilized. The correlation time is proportional to the effective protein volume dependent on protein structure dynamic stability in a certain temperature range which can thus be determined. Sharp temperature changes in the effective volume and their dependence on the concentration of NaCl, ammonium sulphate, polyethylene glycol, and heavy water are interpreted as interconversions of stable low and hightemperature protein conformers with the participation of intermediates. Few variants of the qualitative phase diagrams of water-salt dispersion of the protein were presented on the basis of the results obtained. The regions have been marked, where thermo and salt induced liquid liquid phase transitions can occur, as well as intermediates and supramolecular structures of the protein can form in their course.