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A. Domínguez-Vidal, M. Saenz-Navajas, M.J. Ayora-Cañada, B. Lendl:
"Detection of Albumin Unfolding Preceding Proteolysis Using Fourier Transform Infrared Spectroscopy and Chemometric Data Analysis";
Analytical Chemistry, 78 (2006), 10; 3257 - 3264.

The hydrolysis of bovine serum albumin with protease K at 60° has been studied by means of IR spectroscopy. Two-dimensional correlation spectroscopy (2DCoS) has been used to study spectral changes in the reaction. The use of the multivariate curve resoln.-alternating least-squares method applied to IR measurements allowed the recovery of pure IR spectra and concn. profiles of the different species involved in the reaction. Special attention was paid to the careful inspection of residuals again using 2DCoS. In this way, a heat-induced unfolding step previous to protein hydrolysis was identified. The IR spectra of the intermediate species showed a more disordered structure than native albumin, the decrease in a-helix conformation being esp. noticeable. The formation of b-sheet aggregates due to heating was detected too.