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A proteomic approach to the identification of new tPA receptors in pancreatic cancer cells
Author(s) -
Roda Oriol,
Chiva Cristina,
Espuña Gemma,
Gabius HansJ.,
Real Francisco X.,
Navarro Pilar,
Andreu David
Publication year - 2006
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.200500376
Subject(s) - annexin a2 , receptor , plasminogen activator , microbiology and biotechnology , cell surface receptor , biology , pancreas , proteomics , chemistry , biochemistry , annexin , cell , endocrinology , gene
We have developed a strategy to identify putative tissue‐type plasminogen activator (tPA)receptors present in pancreatic cancer cells by affinity capture with tPA‐Sepharose followed by 2‐DE and MALDI‐MS PMF. Proteins pulled down from either total lysates or raft membrane fractions were characterized and compared with those from a total lysate of an endothelial cell line (HUVEC) to identify pancreas‐restricted tPA receptors. A total of 31 proteins were found by this approach, including annexin A2, already described as a tPA receptor in pancreas and endothelial cells, other proteins acting as tPA receptors ( i.e. , enolase, cytokeratins 8 and 18) in other tissues, and additional proteins not previously identified as candidate tPA receptors. Confirmation of the results was performed for some of these proteins using immunoblotting. These studies are the basis for further functional analyses on the role of these proteins in the biological effects of tPA.