The interaction of gamma-crystallins with model surfaces.

Three biophysical techniques were employed to study the structure and thermal stability of a series of homologous bovine lens gamma-crystallins upon binding to three model surfaces. The surfaces in order of increasing hydrophobicity were silica, methyl silica, and diphenyl silica. Secondary structure was analyzed by deconvolution Fourier transform infrared spectroscopy, while tertiary structure alterations were probed by front surface fluorescence spectroscopy. The effect of surface binding on protein thermal stability was analyzed by fluorescence and differential scanning calorimetry. The comparison of free and surface-bound protein with variations in the electrostatic and hydrophobic character of both the protein and the adsorbent surface with these techniques demonstrated that: (i) destabilization on hydrophobic surfaces is greater than on a more hydrophilic interface, (ii) detectable conformational changes tend to increase as the hydrophobicity of the surface increases, and (iii) subtle structural differences among proteins can play an important role in determining differences in protein stability and structure upon surface adsorption.