Formation mechanism of α-lactalabumin/oleic acid complex characterized by 2D correlation analysis

Partially unfolded α-lactalbumin (ALA) forms a complex with oleic acid (OA) that exhibits cytotoxic activity. In this study, for the first time, the pH-induced formation mechanism for ALA/OA complexes with two different molar ratios was investigated at the molecular level. For a deeper understanding of the formation mechanism of the two different ALA/OA complexes with decreasing pH, principal component analysis (PCA) and two-dimensional (2D) correlation spectroscopy were used to examine the pH-dependent IR spectra of ALA/OA complexes. By tracking the secondary structural variations in the ALA/OA complexes with decreasing pH, we successfully elucidated the formation mechanism of the ALA/OA complexes at the molecular level. The results showed that the secondary structures of theses complexes exhibited the greatest change between pH4 and pH3.5 and that the components that mainly contributed to the pH-induced transition from the N-state to the A-state were dissimilar in the two different ALA/OA complexes.