title: Apolipoprotein C-II amyloid fibrils assemble via a reversible pathway that includes fibril breaking and rejoining.
abstract: Alzheimer's and several other diseases are characterized by the misfolding and assembly of protein subunits into amyloid fibrils. Current models propose that amyloid fibril formation proceeds via the self-association of several monomers to form a nucleus, which then elongates by the addition of monomer to form mature fibrils. We have examined the concentration-dependent kinetics of apolipoprotein C-II amyloid fibril formation and correlated this with the final size distribution of the fibrils determined by sedimentation velocity experiments. In contrast to predictions of the nucleation-elongation model, the final size distribution of the fibrils was found to be relatively independent of the starting monomer concentration. To explain these results, we extended the nucleation-elongation model to include fibril breaking and rejoining as integral parts of the amyloid fibril assembly mechanism. The system was examined under conditions that affected the stability of the mature fibrils including the effect of dilution on the free pool of monomeric apolipoprotein C-II and the time-dependent recovery of fibril size following sonication. Antibody-labelling transmission electron microscopy studies provided direct evidence for spontaneous fibril breaking and rejoining. These studies establish the importance of breaking and rejoining in amyloid fibril formation and identify prospective new therapeutic targets in the assembly pathway.
authors.affiliation: Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, 30 Flemington Road, Parkville, Victoria 3010, Australia.
authors.forename: Katrina J
authors.initials: KJ
authors.lastname: Binger
authors.forename: Chi L L
authors.initials: CL
authors.lastname: Pham
authors.forename: Leanne M
authors.initials: LM
authors.lastname: Wilson
authors.forename: Michael F
authors.initials: MF
authors.lastname: Bailey
authors.forename: Lynne J
authors.initials: LJ
authors.lastname: Lawrence
authors.forename: Peter
authors.initials: P
authors.lastname: Schuck
authors.forename: Geoffrey J
authors.initials: GJ
authors.lastname: Howlett
mesh_terms: D000682:Amyloid / Q000378:metabolism* / Q000648:ultrastructure; D053304:Apolipoprotein C-II / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron; D008958:Models, Molecular; D010316:Particle Size; D020836:Protein Structure, Quaternary; D013010:Sonication
chemical_list: D000682:Amyloid; D053304:Apolipoprotein C-II