Ji H, Reid G E, Moritz R L, Eddes J S, Burgess A W, Simpson R J
Joint Protein Structure Laboratory, Ludwig Institute for Cancer Research (Melbourne Branch), Parkville, Victoria, Australia.
Electrophoresis. 1997 Mar-Apr;18(3-4):605-13. doi: 10.1002/elps.1150180344.
The master two-dimensional gel database of human colon carcinoma cells currently lists cellular proteins from normal crypts and the colorectal cancer cell lines LIM 1863, LIM 1215 and LIM 1899 (Ward et al., Electrophoresis 1990, 11, 883-891; Ji et al., Electrophoresis 1994, 15, 391-405). Updated two-dimensional electrophoretic (2-DE) maps of cellular proteins from LIM 1215 cells, acquired under both nonreducing and reducing conditions, are presented. Fifteen cellular proteins are identified in the reducing 2-DE gel map, and seven in the nonreducing gel map, along with a tabular listing of their M(r)/pI loci and mode of identification. We also include our mass spectrometric based procedures for identifying 2-DE resolved proteins. This procedure relies on a combination of capillary column (0.10-0.32 mm internal diameter) reversed-phase HPLC peptide mapping of in-gel digested proteins, peptide mass fingerprinting, sequence analysis by either collision-induced dissociation or post-source-decay fragmentation, and protein identification using available database search algorithms. These data, and descriptions of the micro-techniques employed in this laboratory for identifying 2-DE resolved proteins can be accessed via the internet URL: http:(/)/www.ludwig.edu.au.
目前，人结肠癌细胞的二维凝胶主数据库列出了来自正常隐窝以及结肠直肠癌细胞系LIM 1863、LIM 1215和LIM 1899的细胞蛋白（沃德等人，《电泳》，1990年，第11卷，883 - 891页；季等人，《电泳》，1994年，第15卷，391 - 405页）。文中展示了在非还原和还原条件下获取的LIM 1215细胞的细胞蛋白的更新二维电泳（2 - DE）图谱。在还原二维凝胶图谱中鉴定出15种细胞蛋白，在非还原凝胶图谱中鉴定出7种，同时列出了它们的相对分子质量/等电点位置及鉴定方式的表格。我们还介绍了基于质谱法鉴定二维电泳分离蛋白的方法。该方法依赖于内径为0.10 - 0.32毫米的毛细管柱反相高效液相色谱对胶内消化蛋白进行肽图谱分析、肽质量指纹图谱分析、通过碰撞诱导解离或源后衰变碎裂进行序列分析以及使用可用数据库搜索算法进行蛋白鉴定。这些数据以及本实验室用于鉴定二维电泳分离蛋白所采用的微技术的描述可通过互联网网址：http://www.ludwig.edu.au获取。
