Surface plasma membrane proteins play important roles in many biological processes such as cell-to-cell interactions, transduction of signals from extracellular factors, and transport of materials.  However, there are challenges in studying these proteins due to the difficulty in preparing clean plasma membrane fractions, their relatively low abundance in comparison to cytosolic proteins, and their hydrophobic nature.  Here, we have combined two cell surface labeling strategies with filter aided sample preparation (FASP) to analyze the cell surface proteome in great depth.  Cell surface capturing (CSC) technology allows for the analysis of N-glycoproteins on the cell surface, whereas cell surface biotinylation (CSB) relies on the use of a membrane-impermeable biotin-conjugated lysine-reactive probe for conjugation with cell surface proteins. Combining these two approaches, we have been able to identify nearly 500 cell surface plasma membrane proteins in various mammalian cell lines.              Arsenite is an environmental contaminant seen in many parts of the world.  Humans have the potential to be exposed to this contaminant through skin contact, inhalation, and ingestion.  In this study, we have employed the aforementioned strategies combined with stable isotope labeling by amino acids in cell culture (SILAC) to discover the aberrant expression of cell surface plasma membrane proteins using a skin fibroblast cell line upon exposure to inorganic arsenite.  Determining which proteins on the cell surface are up and down regulated after exposure to arsenic may allow for a better understanding about the mechanisms of toxicity of this harmful environmental toxicant.

Quantitative analysis of cell surface plasma membrane proteins of skin fibroblasts cells upon arsenite exposure

248th National Meeting (2014)

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