Glycation as a Tool To Probe the Mechanism of β-Lactoglobulin Nanofibril Self-Assembly.

In this study we investigated the effects of different levels of glucosylation and lactosylation on β-Lg self-assembly into nanofibrils at 80 °C and pH 2. Fibrils in heated samples were detected with the thioflavin T assay and transmission electron microscopy, while SDS-PAGE was used to investigate the composition of the heated solutions and fibrils. Glycation had different effects in the nucleation and growth phases. The effect of glycation on the nucleation phase depended on the degree of glycation but not the sugar type, whereas both the type of sugar and the degree of glycation affected the rate of fibril growth. Glycation by either sugar strongly inhibited self-assembly in the growth phase, and lactosylation produced a much stronger effect than glucosylation. We suggest that the varying glycation susceptibility of different lysine residues can explain these observations. The large, polar sugar residues on the glycated fibrillogenic peptides may inhibit fibril assembly by imposing steric restrictions and disrupting hydrophobic interactions.