Figure 5.

Cross-seeding of monomeric filaments. Seeds of monomeric filaments were generated at pH 3, separated from monomers, dialyzed, and resolubilized at pH 7. After concentrating seeds to 20 mg/mL, they were diluted 10-fold into either 20 mg/mL of native lysozyme or buffer only (pH 7, 40 mM NaCl) and incubated at 37 °C. Monomeric filaments (MF) did not induce growth of hew-L monomers as indicated by scattering intensity (a) or ThT fluorescence emission (b). In contrast, protofibrils (PF) generated at pH 7, concentrated (20 mg/mL), and diluted 10-fold into fresh monomers readily induced their self-replication (a). Protofibrils generated at pH 3 could not be used as seeds since they precipitated irreversibly following transfer to pH 7.