A 2-DIMENSIONAL GEL DATABASE OF HUMAN COLON-CARCINOMA PROTEINS

The master two-dimensional gel database of human colon carcinoma cells currently lists cellular proteins from normal crypts and the colorect al cancer cell lines LIM 1863, LIM 1215 and LIM 1899 (Ward et al., Ele ctrophoresis 1990, 11, 883-891; Ji et al., Electrophoresis 1994, 15, 3 91-405). Updated two-dimensional electrophoretic (2-DE) maps of cellul ar proteins from LIM 1215 cells, acquired under both nonreducing and r educing conditions, are presented. Fifteen cellular proteins are ident ified in the reducing 2-DE gel map, and seven in the nonreducing gel m ap, along with a tabular listing of their M-r/pI loci and mode of iden tification. We also include our mass spectrometric based procedures fo r identifying 2-DE resolved proteins. This procedure relies on a combi nation of capillary column (0.10-0.32 mm internal diameter) reversed-p hase HPLC pep tide mapping of in-gel digested proteins, peptide mass f ingerprinting, sequence analysis by either collision-induced dissociat ion or post-source-decay fragmentation, and protein identification usi ng 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.