Chiral and Structural Polymorphism of Fibril Architectures of Homologous Lysozymes

Amyloid fibrils are fascinating and complex structures with the multilayered chiral organization. Using the multimodal methodology, including VCD, ECD, cryo‐EM, and TEM, we characterized in detail different levels of organization (secondary structure/protofilament/mesoscopic structure) of amyloid fibrils prepared from proteins highly homologous in the structure (hen egg white and human lysozymes). Our results demonstrate that small changes in the native protein structure or preparation conditions translate into significant differences in the handedness and architecture of the formed fibrils at various levels of their complexity. In particular, fibrils of hen egg white and human lysozymes obtained in vitro at the same preparation conditions, possess different secondary structure, protofilament twist and ultrastructure. Yet, formed fibrils adopted a relatively similar mesoscopic structure, as observed in high‐resolution 3D cryo‐EM, scarcely used up to now for fibrils obtained in vitro in denaturing condition. Our results add to other puzzling experiments implicating the indeterministic nature of fibril formation. A multimodal approach involving chiroptical spectroscopies (VCD, ECD) and microscopic methods, in particular cryo‐EM, was used to characterize chirality and structure of lysozyme amyloid fibrils. Rich polymorphism was observed for fibrils of hen egg white and human lysozymes with variations in the secondary structures, protofilament handedness and mesoscopic structures.