大鼠嗜铬细胞瘤PC12细胞的蛋白质组学分析。

Proteomic analysis of rat pheochromocytoma PC12 cells.

作者信息

Yang Wei, Liu Peng, Liu Yashu, Wang Qingsong, Tong Yuanpeng, Ji Jianguo

机构信息

Department of Biochemistry and Molecular Biology, College of Life Sciences, National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing, P. R. China.

出版信息

Proteomics. 2006 May;6(10):2982-90. doi: 10.1002/pmic.200500036.

DOI:10.1002/pmic.200500036

PMID:16622837

Abstract

PC12 cell line is well documented and widely applied as many kinds of models in neurobiological and neurochemical studies. Yet a thorough proteomic analysis has not been performed so far. Here we report the construction of a large-scale 2-D protein database for PC12 cells. The proteins extracted from PC12 cells were separated by 2-DE and identified by MALDI-TOF/TOF MS. A total of 1080 protein spots, excised from three different 2-D gels, were identified with high confidence. These proteins represent 474 different gene products, mainly binding proteins and enzymes. Three hundred and seven identified protein spots were located in the low-molecular weight region below 20 kDa. This database today represents one of the largest 2-D databases for higher eukaryotic cell proteomes and for low-molecular weight proteins. In addition, fragment ion spectra obtained by TOF/TOF confirmed that calcylin in PC12 cells was N-acetylated. The database of PC12 proteome is expected to be a powerful tool for neuroscientists.

摘要

PC12细胞系在神经生物学和神经化学研究中作为多种模型已有充分的文献记载且被广泛应用。然而，迄今为止尚未进行全面的蛋白质组学分析。在此，我们报告了一个用于PC12细胞的大规模二维蛋白质数据库的构建。从PC12细胞中提取的蛋白质通过二维电泳进行分离，并通过基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF/TOF MS）进行鉴定。从三块不同的二维凝胶上切下的总共1080个蛋白质点被高可信度地鉴定出来。这些蛋白质代表474种不同的基因产物，主要是结合蛋白和酶。307个已鉴定的蛋白质点位于20 kDa以下的低分子量区域。该数据库目前是高等真核细胞蛋白质组和低分子量蛋白质的最大二维数据库之一。此外，通过飞行时间串联质谱获得的碎片离子光谱证实PC12细胞中的钙调蛋白是N-乙酰化的。PC12蛋白质组数据库有望成为神经科学家的有力工具。