Abstract
Purpose To investigate the role of chaperone activity of α-crystallin in selenite-induced cataract formation.
Methods Selenite cataract was induced in Sprague–Dawley rats by five subcutaneous injections of sodium selenite over a 20-day period starting at 8–10 days postpartum. α-Crystallin was separated from the rat lenses by size-exclusion chromatography. Bovine αL-crystallin and βL-crystallin were isolated for studies in vitro, and for the chaperone assays. The protective effects of both αH- and αL-crystallin were measured spectrophotometrically in four different assay procedures including the thermally induced aggregation of catalase and βL-crystallin, and the fructation- and heat-induced inactivation of catalase. The bovine αL-crystallin was incubated with different concentrations of sodium selenite for 72 h and then its chaperone activity against heat-induced βL-crystallin aggregation was assayed. The aggregation of selenite-treated αL-crystallin was analysed by molecular sieve high-performance liquid chromatography (HPLC).
Results The protection of αH-crystallin was less than that of αL-crystallin in both normal and cataractous lenses. The chaperone activities of both αH- and αL-crystallin in selenite cataract were decreased compared with normal lenses. The protection provided by both αH-crystallin and αL-crystallin against the thermal aggregation of catalase was much greater than their protection against thermally and chemically induced inactivation. HPLC analysis demonstrated aggregation of α-crystallin by sodium selenite after 24 h incubation in a dose-dependent fashion.
Conclusion The chaperone activity of α-crystallin presented parallel patterns of activity with different methods, further supporting the view that the different assays measure essentially the same property. The decreased chaperone activity of α-crystallin in selenite cataract may result from selenite-induced aggregation.
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Acknowledgements
The studies were presented orally at the annual meeting of the European Association for Vision and Eye Research (EVER), Alicante, Spain, October 2001. The authors would like to acknowledge Akbar Z Shyadehi and Barry K Derham for their guidance with the HPLC analysis and aggregation assay. The studies were supported in part by the Wellcome Trust.
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Yan, H., Harding, J., Hui, Y. et al. Decreased chaperone activity of α-crystallin in selenite cataract may result from selenite-induced aggregation. Eye 17, 637–645 (2003). https://doi.org/10.1038/sj.eye.6700419
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DOI: https://doi.org/10.1038/sj.eye.6700419
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