蛋白质组学技术鉴定的PC12细胞的细胞骨架蛋白

The identification of evtoskeleton proteins of PC12 cells with the Broteomic techniaue

目的:使用蛋白质组学的技术手段,鉴定大鼠肾上腺皮质细胞瘤细胞系PC12细胞内的细胞骨架蛋白。方法:提取PC12细胞的蛋白质,建立固相pH梯度双向电泳图谱,应用图像扫描仪及ImageMaster 2D Elite分析软件获得蛋白质点的数字化和匹配性信息,挑选匹配良好的高峰度蛋白点,进行基质辅助激光解吸／电离飞行时间质谱(MALDI- TOF-MS)分析,鉴定。结果:用二维电泳技术分离,并用MALDI-TOF-MS成功鉴定出5个PC12细胞的细胞骨架蛋白。结论:PC12细胞蛋白质组中部分细胞骨架蛋白胶图位点的建立,为今后探讨这一类蛋白在神经系统疾病中的作用奠定了基础,并提供了新的侯选治疗靶点。

Objective：To identify cytoskeleton proteins of PC12 cells with the proteomic technique. Methods： Two-dimentsional polyacrylamide gel electrophoresis （2-DE） map of the proteins extracted from PC12 cells was established by the immobilized pHgradient（IPG） two-dimensional electrophoresis. Protein spot detection and matching were performed by the scanner and the ImageMaster 2D Elite ananlysis software. With the help of Ettanspotter and digester, proteins spots matched well aross the gels were extracted, digested and further identified with assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF MS）. Results： 5 cytoskeleton proteins were separated by 2-DE and identified with MALDI-TOF MS successfully. Conclusions： The establishment of part cytoskeleton proteins map of PC12 cells with the proteomic technique lays foundation for the research on the involvement of cytoskeleton protein in neural disease pathogenesis and offers new therapy targets for them.