Pressure-induced change in proteins studied through chemical modifications.

Pressure-induced change of two bovine proteins, alpha-lactalbumin (LA) and beta-lactoglobulin (LG), was investigated at neutral pH by means of fluorescence and CD spectroscopy. The rate and the extent of modification was considerably increased by applying high pressure during the dansylation reaction of LG, while those for LA were only moderately affected. This difference was accounted for by the structural deformation of these proteins under high pressure. The fluorescence spectrum of these proteins measured under elevated pressure, as well as their fluorescence and CD spectra after the pressure release, indicated different responses towards pressure. The structural change of LA was practically reversible up to 400 MPa, whereas that of LG lost reversibility at 150 MPa or lower. Fluorescent measurement of dansylated (prepared at atmospheric pressure) proteins, especially the energy transfer from the intrinsic Trp residue to the dansyl group, showed that the protein structure was deformed by pressure and that the energy transfer facility of the two proteins was differently affected by high pressure, probably reflecting the degree of compactness of their pressure-perturbed structures.