Effect of protein thermo aggregation on the binding of BSA to gelatin type A

Abstract

We study the effect of limited heat-induced aggregation of BSA on structure development in the water–gelatin-thermally aggregated BSA (BSATA) system. The pH is set at 5.4 and the temperature is higher than the conformation transition temperature of gelatin, but lower than the denaturation temperature of BSA. Dynamic light scattering, circular dichroism, and fluorescence measurements are used to monitor structure changes. Interaction of gelatin with BSATA leads to formation of large complex particles with an average radius ∼1500nm. BSA–gelatin complex formation accompanies partial destabilization of the secondary and tertiary structures of BSA and an additional exposure of hydrophobic tryptophan residues on the surface of the globule. It is shown that electrostatic interaction of the oppositely charged groups of BSATA and gelatin is responsible for formation of such complex particles, whereas the secondary forces (hydrophobic interaction and hydrogen bonds) play an important role in stabilization of the complex particles. The zeta potentials of the native and the thermally aggregated BSA samples were determined, and the solvent quality has been quantified by determining the activity of the protein samples in their saturated solutions. It was shown that steric reasons (large size of the thermally aggregated BSATA particles), and uncomplete charge compensation of the positively charged gelatin molecules by the negatively charged BSATA particles are the main factors in determining structure formation, while the levels of the activity of the native BSA and BSATA have a smaller effect on the structure of complex.