Chaperone Protein Characterization With Multi-Angle Light Scattering

Oligomerization of αB-crystallin is essential for its function as a molecular chaperone. The objective of this study was to delete residues 54FLRAPSWF61 in the N-terminal domain in αB-crystallin and see the effect of deletion on oligomerization and chaperone like function of the protein.

The oligomeric size and hydrodynamic radius of purified recombinant wt-αB and αBD54-61 proteins were determined using a multi-angle dynamic light scattering instrument (DAWN EOS + QELS, Wyatt Technology) coupled to an HPLC fitted with TSKG5000PWXL gel filtration column. The data were acquired and analyzed using ASTRA software. The chaperone activity of the wt-aB and deletion mutant was compared at 37°C using alcohol dehydrogenase (ADH) substrate.