Intermolecular interactions and molecular dynamics in bovine serum albumin solutions studied by small angle X-ray scattering and dielectric relaxation spectroscopy

We used small angle X-ray scattering (SAXS) and dielectric relaxation spectroscopy (DRS) to clarify static structure and molecular dynamics of bovine serum albumin (BSA) in solution. SAXS allowed us to access spatial correlations of the protein molecules in concentrated solutions at different ionic strength. Using a well-established Fourier inversion technique, called SQ-IFT, we deduced the effective pair correlation functions, g(r)(eff), from the effective (or experimental) structure factors, S(q)(eff), without assuming any potential models for protein-protein interactions. In terms of S(q)(eff) and g(r)(eff), the mean nearest-neighbor distance, d*, the osmotic compressibility, kappa(osm), and the coordination number, N-C were evaluated. The complex dielectric spectra of aqueous BSA solutions measured in the frequency range from 1 MHz to 10 GHz were able to be decomposed into three relaxation processes having different physical origins. The highest frequency process, whose relaxation time was ca. 8 ps, was assigned to cooperative rearrangement of H-bond network of bulk-water. We evaluated the effective hydration number of a BSA molecule, Z(hyd), using the bulk-water relaxation amplitude. This approach yielded ca. 1200 hydrated water molecules per a BSA molecule, corresponding to ca. 03 g hydrated water per 1 g protein. The lowest frequency process having a relaxation time of ca. 50 ns was assigned to rotational diffusion of the BSA molecule. We confirmed that an effective molar volume of BSA calculated with Stokes-Einstein-Debye equation was identical to that predicted from its molecular weight and specific density. According to the Kirkwood relationship, the dipole-dipole orientational correlation factor decreased with an increase of protein concentration, which indicated the operation of BSA-BSA interaction and favored antiparallel dipolar correlation between the protein molecules. (C) 2014 Elsevier B.V. All rights reserved.