Cross-β-Sheet Structure in Amyloid Fiber Formation

As proteins aggregate to form amyloid fibers, their secondary structure changes from its native form to cross-β-sheet. Whether this conformational change is essential for fiber formation remains unknown. Evidence from atomic force microscopy and transmission electron microscopy suggests that aggregation occurs in two stages. Initially, protein monomers aggregate into colloidal spheres; however, they stop growing after reaching a uniform diameter. The spheres then join together to form linear chains which evolve into mature fibers. In this paper, we apply, for the first time, the DLVO theory, formulated by Derjaguin, Landau, Verwey and Overbeek for the quantitative analysis of colloidal interactions, to elucidate the two stages of fiber formation. We find that, as like-charged protein molecules aggregate, the total charge of the colloidal sphere increases until it repels additional monomers from coming close enough to bind, limiting the size of the colloidal particle. Energy analysis and X-ray diffraction data suggest that aggregation of multiple protein monomers onto the growing colloid drives their misfolding into hairpin loops. These loops stack together to form a U-shaped trough which initially adopts a cross-α-sheet structure with a strong dipole moment. Driven by charge−dipole interactions, the colloidal spheres aggregate into a linear chain. The peptide strands are oriented perpendicular to the direction of the dipole of each sphere and, therefore, are also perpendicular to the axis of the linear chain as it forms and evolves into the mature fiber. The cross α-sheet then evolves into the thermodynamically more stable cross β-sheet.