Early stages of misfolding and association of beta2-microglobulin: insights from infrared spectroscopy and dynamic light scattering.
Conformational changes associated with the assembly of recombinant beta 2-microglobulin in vitro under acidic conditions were investigated using infrared spectroscopy and static and dynamic light scattering. In parallel, the morphology of the different aggregated species obtained under defined conditions was characterized by electron microscopy. The initial salt-induced aggregate form of beta 2-microglobulin, composed of small oligomers (dimers to tetramers), revealed the presence of beta-strands organized in an intramolecular-like fashion. Further particle growth was accompanied by the formation of intermolecular beta-sheet structure and led to short curved forms. An increase in temperature by only 25 degrees C was able to disaggregate these assemblies, followed by the formation of longer filamentous structures. In contrast, a rise in temperature up to 100 degrees C was associated with a reorganization of the short curved forms at the level of secondary structure and the state of assembly, leading to a species with a characteristic infrared spectrum different from those of all the other aggregates observed before, suggesting a unique overall structure. The infrared spectral features of this species were nearly identical to those of beta 2-microglobulin assemblies formed at low ionic strength with agitation, indicating the presence of fibrils, which was confirmed by electron microscopy. The observed spectroscopic changes suggest that the heat-triggered conversion of the short curved assemblies into fibrils involves a reorganization of the beta-strands from an antiparallel arrangement to a parallel arrangement, with the latter being characteristic of amyloid fibrils of beta 2-microglobulin.
Fabian
Heinz
H
Robert Koch-Institut, P 25 and ZBS4, Nordufer 20, D-13353 Berlin, Germany. fabianh@rki.de
Gast
Klaus
K
Laue
Michael
M
Misselwitz
Rolf
R
Uchanska-Ziegler
Barbara
B
Ziegler
Andreas
A
Naumann
Dieter
D
D000682:Amyloid / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008027:Light; D008854:Microscopy, Electron; D017510:Protein Folding; D013055:Spectrophotometry, Infrared; D013696:Temperature; D001613:beta 2-Microglobulin / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure
D000682:Amyloid; D001613:beta 2-Microglobulin