应用蛋白质组学方法分析猪晶状体不同区域上皮细胞中蛋白质的表达差异

The Proteomics Analysis of Different Expression between the Central and the Peripheral Epithelial Cells in Porcine Lens

应用蛋白质组学方法分析比较猪晶状体中央和周边上皮细胞的蛋白质表达差异。将32个正常猪晶状体前囊膜所附着于的上皮细胞分为中央和周边两部分,经二维凝胶电泳分离和凝胶考马斯亮兰染色,质谱(MALD I-TOF-MS)鉴定差异蛋白质斑点,并对鉴定的蛋白质进行分类。结果显示来自中央与周边区域的猪晶体上皮细胞的蛋白在二维凝胶上分别有801和886个蛋白质斑点,鉴定出差异表达蛋白84个。差异表达的蛋白质在功能上有一定趋向性,主要涉及代谢、细胞骨架、信号转导/细胞周期/转录因子等。

The difference of the protein expressions between central and peripheral epithelial cells of porcine lens was investigated using proteomics techniques. The anterior sub-capsular epithelial cells adherent to the lens capsule from 32 porcine lenses were dissected into central（6 mm in diameter） and peripheral parts. The extracted proteins from central and peripheral epithelial cells were isolated by two-dimensional gel electrophoresis（IEF/SDS- PAGE, 2D） with Coomassie blue staining. Representative gel patterns were compared by software. Differential spots observed on two 2D gels were selected and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry （MALDI - TOF - MS）. Overall, 801 and 886 spots were detected on the 2D gel of the central and peripheral sampies, respectively. Eighty-four spots were identified by MS, which appeared to be present or to be intensive on one gel, but were absent or weak on another one. Most proteins expressed differently can be classified, based on their functions, into metabolism, cytoskeleton, signaling/cell cycle/transcription and others.