Ortwerth B J, Olesen P R, Sharma K K
Exp Eye Res. 1986 Dec;43(6):955-63. doi: 10.1016/0014-4835(86)90073-4.
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.
据报道，有一种方法可通过在低盐缓冲液中对悬浮的蛋白质进行短时间超声处理来实现晶状体大部分水不溶性部分的增溶。该程序可溶解超过90%的牛晶状体水不溶性部分和80%的正常人晶状体水不溶性部分。从白内障晶状体中溶解的蛋白质减少，但在每种情况下，超声处理至少相当于用6.0 M尿素提取。通过琼脂糖A-1.5 m凝胶过滤色谱对溶解的蛋白质进行分级分离，显示牛晶状体的天然分子量，但人晶状体提取物仅部分解聚。通过超声处理和6.0 M尿素提取溶解的蛋白质的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）比较显示，除了超声处理溶解了更多留在凝胶顶部的高度交联蛋白质外，没有重大差异。
