Competing pathways determine fibril morphology in the self-assembly of beta2-microglobulin into amyloid.
Despite its importance in biological phenomena, a comprehensive understanding of the mechanism of amyloid formation remains elusive. Here, we use atomic force microscopy to map the formation of beta2-microglobulin amyloid fibrils with distinct morphologies and persistence lengths, when protein concentration, pH and ionic strength are varied. Using the resulting state-diagrams, we demonstrate the existence of two distinct competitive pathways of assembly, which define an energy landscape that rationalises the sensitivity of fibril morphology on the solution conditions. Importantly, we show that semi-flexible (worm-like) fibrils, which form rapidly during assembly, are kinetically trapped species, formed via a non-nucleated pathway that is explicitly distinct from that leading to the formation of the relatively rigid long-straight fibrils classically associated with amyloid. These semi-flexible fibrils also share an antibody epitope common to other protein oligomers that are known to be toxic species linked to human disease. The results demonstrate the heterogeneity of amyloid assembly, and have important implications for our understanding of the importance of oligomeric states in amyloid disease, the origins of prion strains, and the development of therapeutic strategies.
Gosal
Walraj S
WS
School of Physics and Astronomy, University of Leeds, Leeds LS2 9JT, UK.
Morten
Isobel J
IJ
Hewitt
Eric W
EW
Smith
D Alastair
DA
Thomson
Neil H
NH
Radford
Sheena E
SE
D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D046912:Multiprotein Complexes; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D001613:beta 2-Microglobulin / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure
D000682:Amyloid; D046912:Multiprotein Complexes; D011994:Recombinant Proteins; D001613:beta 2-Microglobulin