The effect of arginine on inhibiting amyloid fibril derived from β-casein and the binding studies with multi-spectroscopic techniques.

In general, β-casein is a stable molecular chaperone. However, the fact that amyloid fibrils derived from β-casein has been reported in some cases, which were usually associated with some malignant breast diseases. As an important amino acid, arginine not only exhibits the significance in casein synthesis in mammary gland, but also has great potentiality in inhibiting the amyloid fibril formation. Therefore, the influence of arginine on the amyloid fibrils formed by β-casein and further molecular mechanism were studied firstly with multi-spectroscopic techniques in the present work. The results of Thioflavin T determination, particle size analysis, transmission electron microscope observation showed that arginine not only inhibited the aggregation of β-casein at the growth stage, but also depolymerized the mature amyloid fibrils at the saturation stage. The further fluorescence experiment results demonstrated that the complex was formed between β-casein and arginine. Besides, there was one binding site and 0.48 nm binding distance. The thermodynamic parameters like ΔG0, ΔS0, ΔH0 were all negative, showing their binding reaction was spontaneous, and hydrogen bond and van der Waals force were the possibly chief intermolecular forces. Furthermore, the synchronous fluorescence spectra showing that the conformation of β-casein was affected and its tyrosine residues were gradually buried inside the protein. Our research would provide new insights into the treatments for the breast amyloidosis.