title: IR spectroscopic analyses of amyloid fibril formation of β2-microglobulin using a simplified procedure for its in vitro generation at neutral pH.
abstract: β2-microglobulin (β2m) is known to be the major component of fibrillar deposits in the joints of patients suffering from dialysis-related amyloidosis. We have developed a simplified procedure to convert monomeric recombinant β2m into amyloid fibrils at physiological pH by a combination of stirring and heating, enabling us to follow conformational changes associated with the assembly by infrared spectroscopy and electron microscopy. Our studies reveal that fibrillogenesis begins with the formation of relatively large aggregates, with secondary structure not significantly altered by the stirring-induced association. In contrast, the conversion of the amorphous aggregates into amyloid fibrils is associated with a profound re-organization at the level of the secondary and tertiary structures, leading to non-native like parallel arrangements of the β-strands in the fully formed amyloid structure of β2m. This study highlights the power of an approach to investigate the formation of β2m fibrils by a combination of biophysical techniques including IR spectroscopy.
authors.affiliation: Robert Koch-Institut, ZBS 6, Nordufer 20, D-13353 Berlin, Germany. fabianh@rki.de
authors.forename: Heinz
authors.initials: H
authors.lastname: Fabian
authors.forename: Klaus
authors.initials: K
authors.lastname: Gast
authors.forename: Michael
authors.initials: M
authors.lastname: Laue
authors.forename: Katharina J
authors.initials: KJ
authors.lastname: Jetzschmann
authors.forename: Dieter
authors.initials: D
authors.lastname: Naumann
authors.forename: Andreas
authors.initials: A
authors.lastname: Ziegler
authors.forename: Barbara
authors.initials: B
authors.lastname: Uchanska-Ziegler
mesh_terms: D000682:Amyloid / Q000138:chemical synthesis* / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D013055:Spectrophotometry, Infrared; D013696:Temperature; D001613:beta 2-Microglobulin / Q000737:chemistry*
chemical_list: D000682:Amyloid; D001613:beta 2-Microglobulin
keywords: Amyloid fibril; Amyloidogenesis; DLS; DSC; EM; IR; IR spectroscopy; LS; SLS; ThT; WL; differential scanning calorimetry; dynamic light scattering; electron microscopy; infrared; long straight; static light scattering; thioflavin T; worm-like; β(2)-microglobulin; β(2)-microglobulin fibrils formed at acidic pH; β(2)-microglobulin fibrils formed at physiological pH; β(2)m; β(2)m-F(Acid); β(2)m-F(Phys)