J Biol Chem 2002 Dec 10; [epub ahead of print]
Comprehensive proteomic analysis of breast cancer cell membranes reveals unique proteins with potential roles in clinical cancer.
Adam PJ, Boyd R, Tyson KL, Fletcher GC, Stamps A, Hudson L, Poyser HR, Redpath N, Griffiths M, Steers G, Harris AL, Patel S, Berry J, Loader JA, Townsend RR, Daviet L, Legrain P, Parekh R, Terrett JA.
Molecular Biology, Oxford Glycosciences, Abingdon, Oxon OX14 4RY.
Proteins associated with cancer cell plasma membranes are rich in known drug and antibody targets as well as other proteins known to play key roles in the abnormal signal transduction processes required for carcinogenesis. We describe here a proteomics process that comprehensively annotates the protein content of breast tumour cell membranes and defines the clinical relevance of such proteins. Tumour-derived cell lines were used to ensure an enrichment for cancer cell specific plasma membrane proteins as it is difficult to purify cancer cells and then obtain good membrane preparations from clinical material. Multiple cell lines with different molecular pathologies were used to represent the clinical heterogeneity of breast cancer. Peptide tandem mass spectra were searched against a comprehensive database containing known and conceptual proteins derived from many public databases including the draft human genome sequences. This plasma membrane enriched proteome analysis created a database of more than 500 breast cancer cell line proteins, 27% of which were of unknown function. The value of our approach is demonstrated by further detailed analyses of three previously uncharacterised proteins whose clinical relevance has been defined by their unique cancer expression profiles and the identification of protein binding partners that elucidate potential functionality in cancer. |
