title: Amyloid fibril formation of the mouse V(L) domain at acidic pH.
abstract: The recombinant V(L) domain that represents the variable part of the light chain (type kappa) of mouse monoclonal antibody F11 directed against human spleen ferritin was found to form amyloid fibrils at acidic pH as evidenced by electron microscopy, thioflavin T binding, and apple-green birefringence after Congo red staining. This is the first demonstration of amyloid fibril formation of the mouse V(L) domain. To understand the mechanism of acidic pH-induced amyloid fibril formation, conformational changes of the V(L) domain were studied by one-dimensional NMR, differential scanning calorimetry, analytical ultracentrifugation, hydrophobic dye binding, far-UV circular dichroism, and tryptophan fluorescence. The results indicated accumulation of two intermediate states during acid unfolding, which might be responsible for amyloid fibril formation. The more structured intermediate that exhibited maximal accumulation at pH 3 retained the nativelike secondary structure and a hydrophobic core, but exposed hydrophobic surfaces that bind 8-anilino-1-naphthalenesulfonate. Below pH 2, a more disordered intermediate with dequenched tryptophan fluorescence but still retaining the beta-sheet structure accumulated. The optimal pH of amyloid fibril formation (i.e., pH 4) was close to the optimal pH of the accumulation of the nativelike intermediate, suggesting that the amyloid fibrils might be formed through this intermediate.
authors.affiliation: Institute of Bio-Organic Chemistry, National Academy of Sciences of Belarus, Minsk 220141, Belarus. martsev@iboch.bas-net.by
authors.forename: Sergey P
authors.initials: SP
authors.lastname: Martsev
authors.forename: Anatoly P
authors.initials: AP
authors.lastname: Dubnovitsky
authors.forename: Alexander P
authors.initials: AP
authors.lastname: Vlasov
authors.forename: Masaru
authors.initials: M
authors.lastname: Hoshino
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Hasegawa
authors.forename: Hironobu
authors.initials: H
authors.lastname: Naiki
authors.forename: Yuji
authors.initials: Y
authors.lastname: Goto
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000648:ultrastructure; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D051379:Mice; D008854:Microscopy, Electron; D019906:Nuclear Magnetic Resonance, Biomolecular; D014364:Tryptophan / Q000737:chemistry
chemical_list: D000682:Amyloid; D007135:Immunoglobulin Variable Region; D014364:Tryptophan