Abstract
The molecular chaperone αB-crystallin is a small heat shock protein that inhibits the aggregation of, among others, Aβ42 and α-synuclein. These proteins are major hallmarks of Alzheimer's and Parkinson's disease, respectively. In order to understand the mechanism with which αB-crystallin performs its chaperone function it is essential to characterize its self-assembly in terms of aggregate size distribution, structure, and critical concentration. The size distribution of the assemblies has been widely discussed and they have been suggested to be monodisperse or polydisperse with varying size distributions covering a range of 10-40 monomers per assembly. Here, the size distribution was studied using dynamic and static light scattering, microfluidic diffusional sizing (MDS), as well as small-angle X-ray scattering (SAXS). Findings indicate that αB-crystallin has a preference toward forming spherical assemblies consisting of 18 monomers with a hydrodynamic radius of ≈7 nm after one week. SAXS data were modelled using a homogeneous sphere model with a radius of 6 nm, which is comparable to the light scattering and MDS results. 2D classes built from negative stain transmission electron microscopy images suggest that the spherical aggregates contain several smaller globular units. Furthermore, the findings show that the size of the assemblies is independent of protein concentration, supporting a strong preference for specific assembly constellations.