title: Role of the N and C-terminal strands of beta 2-microglobulin in amyloid formation at neutral pH.
abstract: Beta 2-microglobulin (beta(2)m) is known to form amyloid fibrils de novo in vitro under acidic conditions (below pH 4.8). Fibril formation at neutral pH, however, has only been observed by deletion of the N-terminal six residues; by the addition of pre-assembled seeds; or in the presence of Cu(2+). Based on these observations, and other structural data, models for fibril formation of beta(2)m have been proposed that involve the fraying of the N and C-terminal beta-strands and the consequent loss of edge strand protective features. Here, we examine the role of the N and C-terminal strands in the initiation of fibrillogenesis of beta(2)m by creating point mutations in strands A and G and comparing the properties of the resulting proteins with variants containing similar mutations elsewhere in the protein. We show that truncation of buried hydrophobic side-chains in strands A and G promotes rapid fibril formation at neutral pH, even in unseeded reactions, and increases the rate of fibril formation under acidic conditions. By contrast, similar mutations created in the remaining seven beta-strands of the native protein have little effect on the rate or pH dependence of fibril formation. The data are consistent with the view that perturbation of the N and C-terminal edge strands is an important feature in the generation of assembly-competent states of beta(2)m.
authors.affiliation: School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Susan
authors.initials: S
authors.lastname: Jones
authors.forename: David P
authors.initials: DP
authors.lastname: Smith
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D009154:Mutation; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D014508:Urea / Q000494:pharmacology; D001613:beta 2-Microglobulin / Q000737:chemistry* / Q000502:physiology*
chemical_list: D000682:Amyloid; D001613:beta 2-Microglobulin; D014508:Urea