RT Journal Article
SR Electronic
T1 Proteomic Analysis of Membrane-associated Proteins from the Breast Cancer Cell Line MCF7
JF Cancer Genomics - Proteomics
JO Cancer Genomics Proteomics
FD International Institute of Anticancer Research
SP 199
OP 207
VO 2
IS 4
A1 C. PIONNEAU
A1 L. CANELLE
A1 J. BOUSQUET
A1 J. HARDOUIN
A1 J. BIGEARD
A1 M. CARON
A1 R. JOUBERT-CARON
YR 2005
UL http://cgp.iiarjournals.org/content/2/4/199.abstract
AB Background: Proteins associated with cancer cell membranes represent targets of choice for humoral immune response as well as potential tumour marker proteins in human malignancies. However, proteomic analysis of these proteins, and more generally of low-soluble proteins, remains difficult. Materials and Methods: The breast cancer cell line MCF7 was selected to evaluate a sequential extraction method that enables simple fractionation of human cell proteins according to their subcellular localization, yielding subproteomes enriched in cytosolic and membrane-associated proteins, respectively. A crude plasma membrane preparation was followed by the solubilisation of proteins using trifluoroethanol (TFE) as co-solvent. Results: Cross-matching and statistical analysis performed for each set of two-dimensional electrophoresis (whole-cell, membrane and soluble extracts) and between the different sets highlighted the reproducibility of the extraction process and its usefulness for proteomic analysis. Eighty-three % of the spots of the gels corresponding to the membrane fraction were not found in the gels of the soluble fraction. Conclusion: Due to its simplicity, the approach described here appears well suited for membrane proteomic investigation of human cancer cells and detection of potential biomarkers undetected by current techniques.