title: A helical structural nucleus is the primary elongating unit of insulin amyloid fibrils.
abstract: Although amyloid fibrillation is generally believed to be a nucleation-dependent process, the nuclei are largely structurally uncharacterized. This is in part due to the inherent experimental challenge associated with structural descriptions of individual components in a dynamic multi-component equilibrium. There are indications that oligomeric aggregated precursors of fibrillation, and not mature fibrils, are the main cause of cytotoxicity in amyloid disease. This further emphasizes the importance of characterizing early fibrillation events. Here we present a kinetic x-ray solution scattering study of insulin fibrillation, revealing three major components: insulin monomers, mature fibrils, and an oligomeric species. Low-resolution three-dimensional structures are determined for the fibril repeating unit and for the oligomer, the latter being a helical unit composed of five to six insulin monomers. This helical oligomer is likely to be a structural nucleus, which accumulates above the supercritical concentration used in our experiments. The growth rate of the fibrils is proportional to the amount of the helical oligomer present in solution, suggesting that these oligomers elongate the fibrils. Hence, the structural nucleus and elongating unit in insulin amyloid fibrillation may be the same structural component above supercritical concentrations. A novel elongation pathway of insulin amyloid fibrils is proposed, based on the shape and size of the fibrillation precursor. The distinct helical oligomer described in this study defines a conceptually new basis of structure-based drug design against amyloid diseases.
authors.affiliation: Department of Medicinal Chemistry, University of Copenhagen, Copenhagen, Denmark. bv@farma.ku.dk
authors.forename: Bente
authors.initials: B
authors.lastname: Vestergaard
authors.forename: Minna
authors.initials: M
authors.lastname: Groenning
authors.forename: Manfred
authors.initials: M
authors.lastname: Roessle
authors.forename: Jette S
authors.initials: JS
authors.lastname: Kastrup
authors.forename: Marco
authors.initials: M
authors.lastname: van de Weert
authors.forename: James M
authors.initials: JM
authors.lastname: Flink
authors.forename: Sven
authors.initials: S
authors.lastname: Frokjaer
authors.forename: Michael
authors.initials: M
authors.lastname: Gajhede
authors.forename: Dmitri I
authors.initials: DI
authors.lastname: Svergun
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D007328:Insulin / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation; D014961:X-Ray Diffraction
chemical_list: D000682:Amyloid; D007328:Insulin