The molecular chaperone function of alpha-crystallin is impaired by UV photolysis.

Buffer solutions of the lens protein gamma-crystallin and the enzymes aldolase and liver alcohol dehydrogenase became turbid and formed solid precipitate upon exposure to an elevated temperature of 63 degrees C or to UV radiation at 308 nm. When alpha-crystallin was added to the protein solutions in stoichiometric amounts, heat or UV irradiation did not cause turbidity, or turbidity developed much less rapidly than in the absence of alpha-crystallin. Hence, normal alpha-crystallin functioned as a "molecular chaperone," providing protection against both UV and heat-induced protein aggregation. When alpha-crystallin was preirradiated with UV at 308 nm, its ability to function as a chaperone vis-a-vis both UV and heat-induced aggregation was significantly impaired, but only at relatively high UV doses. A major effect of preirradiation of alpha-crystallin was to cause interpeptide crosslinking among the alpha A2 and alpha B2 subunits of the alpha-crystallin macromolecule. In our experiments alpha-crystallin was exposed to UV doses, which resulted in 0.50 and 90% crosslinking as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. alpha-Crystallin samples that were 50% and 90% crosslinked gave chaperone protection, which was increasingly impaired relative to unirradiated alpha-crystallin. The results are consistent with the notion that UV irradiation of alpha-crystallin results in loss of chaperone binding sites.