遗传性白内障小鼠晶状体的比较蛋白质组学分析

Objective To separate total lens proteins of congenital inherited cataract in mice and observe the alteration of proteins after gene mutation. Methods We studied the mice with a spontaneous mutation of crystallin gamma S (Crygs) transmitted as a recessive trait. Total proteins were extracted and separated using immobilized pH gradient (IPG), two-dimensional electrophoresis (2-DE) and colloidal Coomassie Brilliant Blue (CBB) staining. And then image analysis was carried out using PDQuest 7.30 software package. Several significantly differential proteins in gel were identified by matrix assisted laser adsorption/ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS). Results As the 882μg sample was added, we detected 417±53 spots and 370±41spots in cataract and normal lens respectively. As the 190μg sample was loaded, we detected 60±7 spots and 57±5 spots in cataract and normal lens respectively. Seven kinds of differential proteins were identified, including BFSP/filensin, γS, γF, βA1, βB1, βB2 andαB. In crystalline lens of mutant mice, γS and beaded-filament structure protein (BFSP/filensin) were not detected. γF was down expressed (<4 fold) whileβA1, βB1, βB2 and αB were over expressed (>4 fold) in mutant cataract. The latter proteins have less MW than normal. It suggests they were possibly truncated. Conclusion 2-DE and mass spectrometry can help to assess and analyze the function of proteins as a novel approach. The mutant Crygs gene can lead to the abnormity of γS crystallin, which can induce the changes of skeletonal protein (BFSP/filensin) and other crystallins (γF, βA1, βB1, βB2 and αB) and then evoke cataract secondarily.