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Subcellular proteomics of cell differentiation: Quantitative analysis of the plasma membrane proteome of Caco‐2 cells
Author(s) -
Pshezhetsky Alexey V.,
Fedjaev Michael,
Ashmarina Lyudmila,
Mazur Alexander,
Budman Lorne,
Sinnett Daniel,
Labuda Damian,
Beaulieu JeanFrançois,
Ménard Daniel,
Nifant'ev Ilya,
Levy Émile
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600956
Subject(s) - proteome , membrane protein , proteomics , biology , transmembrane protein , microbiology and biotechnology , biochemistry , membrane , cell , chemistry , gene , receptor
Human colorectal carcinoma (Caco‐2) cells undergo in culture spontaneous enterocytic differentiation, characterized by polarization and appearance of the functional apical brush border membrane. To provide insights into the biology of differentiation, we have performed a comparative proteomic analysis of the plasma membranes from proliferating cells (PCs) and the apical membranes from differentiated cells (DCs). Proteins were resolved by SDS‐PAGE, in‐gel digested and analyzed by RP‐LC and MS/MS. Alternatively, proteins were digested in solution, and tryptic peptides were labeled with isotopic tags and analyzed by 2‐D LC followed by MS/MS. Among the 1125 proteins identified in both proteomes, 76 were found to be significantly increased in the membranes of DCs and 61 were increased in PCs. Majority of the proteins increased in the apical membranes were metabolic enzymes, proteins involved in the maintenance of cellular structure, transmembrane transporters, and proteins regulating vesicular transport. In contrast, majority of the proteins increased in the membranes of PCs were involved in gene expression, protein synthesis, and folding. Both groups contained many novel proteins with yet to be identified functions, which could provide potential new markers of the intestinal cells or of colorectal cancer.