title: Reduction of disulfide bonds in an amyloidogenic Bence Jones protein leads to formation of "amyloid-like" fibrils in vitro.
abstract: Motivated by the finding that the amino acid sequence of the Bence Jones protein BJP-DIA was identical to that of the main protein component of the amyloid fibrils obtained from the same patient with AL-amyloidosis, (Klafki, H.-W., Kratzin, H.-D., Pick, A.-I., Eckart, K., Karas, M. & Hilschmann, N. (1992) Biochemistry 31, 3265-3272.), we attempted to create "amyloid-like" fibrils from the Bence Jones protein in vitro, without addition of proteolytic enzymes. Reduction of BJP-DIA, solubilized in PBS, pH 7.4, overnight at 37 degrees C resulted in the formation of a precipitate which had affinity for the dye Congo red. Electron microscopy of negatively stained samples of the reduced protein revealed aggregates of linear unbranched fibrils. SDS-polyacrylamide gel electrophoresis demonstrated that the precipitate consisted almost exclusively of intact light chain molecules. This result makes it possible to deduce a molecular model of these amyloid fibrils generated in vitro.
authors.affiliation: Max-Planck-Institute for Experimental Medicine, Göttingen, Germany.
authors.forename: H W
authors.initials: HW
authors.lastname: Klafki
authors.forename: A I
authors.initials: AI
authors.lastname: Pick
authors.forename: I
authors.initials: I
authors.lastname: Pardowitz
authors.forename: T
authors.initials: T
authors.lastname: Cole
authors.forename: L A
authors.initials: LA
authors.lastname: Awni
authors.forename: H U
authors.initials: HU
authors.lastname: Barnikol
authors.forename: F
authors.initials: F
authors.lastname: Mayer
authors.forename: H D
authors.initials: HD
authors.lastname: Kratzin
authors.forename: N
authors.initials: N
authors.lastname: Hilschmann
mesh_terms: D000682:Amyloid / Q000096:biosynthesis*; D001536:Bence Jones Protein / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D002852:Chromatography, Ion Exchange; D004220:Disulfides / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008854:Microscopy, Electron; D010084:Oxidation-Reduction
chemical_list: D000682:Amyloid; D004220:Disulfides; D001536:Bence Jones Protein