用蛋白质印迹转移技术分析正常及硒性白内障大鼠晶状体及房水中蛋白质的性质

本文用蛋白质印迹转移技术分析了正常及硒性白内障大鼠晶状体及房水中蛋白质的性质。结果表明,晶状体中的脲溶性蛋白质可被抗α及抗γ晶体蛋白血清识别,提示α及γ晶体蛋白均为脲溶性蛋白质的主要成份。患白内障时房水中的蛋白质含量明显增加,且主要被抗γ血清识别,而被抗α血清识别的成份很少,表明在大鼠硒性白内障形成过程中,有较多低分子量蛋白质漏出到房水中,且其主要成份为γ晶体蛋白。此外,我们还发现正常及硒性白内障大鼠晶状体膜蛋白质与抗α及抗γ血清起反应的程度及分布有所不同,提示晶状体细胞膜与晶体蛋白之间存在着相互作用。

Western Blot Analysis of Lens Proteins and Aqueous Humor in Normal and Cataractous Rat Eyes

Aqueous humor was collected,and water-soluble,urea-soluble and membrane proteins in lenses were isolated from both normal and selenium induced catarac-tous rat eyes.Confirmation for the presence of both a and Y crystalline in urea-sol- uble proteins was demonstrated by Western tmmunoblotting with antibodies against **********a and ***********r crystallins.Increase of proteins in the aqueous humor from rats with cataract was observed.Immunoblots revealed that the major components in aqueous humor could be stained with anti-**********r crystallin but only a few of them were stained with anti-**********a cry-stallin.These results demonstrate that increased urea-soluble proteins in the cataractous lenses were derived mainly from the aggregation of a and *********r crystallins,along with the selective leakage of **********r crystallin into the aqueous humor could account for the decrease in water-soluble proteins in lens during cataract formation.We also found some difference in membrane proteins between normal and cataractous lenses by imm-unoblots staining with anti-************a and anti-*********r crystallins,suggesting that significant interactions do exist between lens membrane proteins and soluble crystallins.