Proteomic Analysis of Human Intestinal Epithelial Cells

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A major challenge in cell biology is to identify the subcellular distribution of proteins within cells, and to characterize how protein localization changes under different cell growth conditions and in response to stress and other external signals. Protein localization is usually determined either by microscopy, or using cell fractionation combined with protein blotting techniques. Both these approaches are intrinsically low throughput and limited to the analysis of known components. Our objective is to establish proteomic based methods to characterize the differences in protein expression and subcellular localisation between different cell lines and extend this characterization to human tissues. These approaches will provide a basis for further proteomic characterisation of corresponding human tissues from normal and diseases to identify potential biomarkers and study normal functions of cells in tissues. More specifically, we will use this method to compare normal epithelial intestinal cells and different cell lines and tissues. Our hypothesis is that protein expression and localization profiling will allow us to identify proteins involved in cell transformation and differentiation specific to these tissues.