Molecular chaperone activity of Alpha-Crystallin via protection of

Aquaporin0 (AQP0) from thermal stress

YaQuesha Culbreth1*, Nhi Le2 and Satyanarayana Swamy Mruthinti3
1-3

Department of Biology, University of West Georgia, Carrollton 30118,

Georgia State, USA
ABSTRACT

-Crystallins are molecular chaperones that protect vertebrate eye lens proteins from
detrimental protein aggregation. The present study is aimed to investigate the
molecular chaperone activity of alpha crystalline via protection of Aquaporin0 (AQP0)
under thermal stress. Lens membranes are prepared from calf eyes. Thermal
denaturation of calf lens AQP0 was performed to initiate aggregation. The extent of
aggregation was measured either in the absence or in the presence of -crystallin. To
characterize the protein aggregation, proteins were separated on a 12% SDSpolyacrylamide gel. The results show an increase in the amount of AQP0 that
aggregated as the temperature increased from 75C to 90C. The results obtained
from SDS-PAGE suggest that AQP0 monomers completely disappear at 75C and
beyond, leading to the formation of large aggregates >200 kDa. -Crystallin,
molecular chaperone abundantly present in the eye lens, completely prevented the
AQP0 aggregation at a 1:1 (weight/weight) ratio. In contrast to the general
observation, -crystallin exhibited better chaperone-like activity towards AQP0. Crystallin offers similar protection against thermal aggregation as in the case of the
non-aggregated AQP0, suggesting that -crystallin may bind to either intracellular
loops or other residues of AQP0 that become exposed during thermal stress. Therefore
this study concludes that the chaperone function of -crystallin works against
membrane proteins such as AQP0, which is involved in maintaining lens fiber cell
homeostasis and lens transparency.
Key words: Aquaporin0, -Crystallins, Chaperone, Thermal stress.

Title:

Protection of Aquaporin0 (AQPO) from thermal stress by Alpha-Crystallin via its

molecular chaperone function