Solubilization of the lens water-insoluble fraction by sonication.
A method is reported whereby the solubilization of the bulk of the lens water-insoluble fraction is accomplished by a short sonication of the suspended proteins in low salt buffers. This procedure solubilized greater than 90% of a bovine lens water-insoluble fraction and 80% of the normal human lens water-insoluble fraction. Decreased protein was solubilized from cataractous lenses, but in every case sonication was at least equivalent to extraction with 6.0 M urea. Fractionation of the solubilized proteins by Agarose A-1.5 m gel filtration chromatography showed native molecular weights for bovine lens, but only partial disaggregation with human lens extracts. A sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) comparison of the proteins solubilized by sonication and 6.0 M urea extraction showed no major differences except that sonication solubilized more of the highly cross-linked protein which remained at the top of the gel.
D000328:Adult; D000368:Aged; D000818:Animals; D002386:Cataract / Q000378:metabolism; D002417:Cattle; D002850:Chromatography, Gel; D003459:Crystallins / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008875:Middle Aged; D012995:Solubility; D013010:Sonication; D013997:Time Factors; D014465:Ultrasonics
D003459:Crystallins