title: Profiling of proteolytic activities secreted by cancer cells using phage display substrate technology.
abstract: Although the cellular steps required for metastasis are similar for all cancer cells, proteases involved in this process and their expression levels vary greatly between different cancer types. Thus, the identification of these proteolytic activities represents a crucial issue in the understanding of cancer development. Until now, phage display substrate technology has been successfully employed for the characterization of purified proteases but was never used with a mix of proteases. In the present work, we report an easy protocol to identify multiple proteolytic activities secreted by cancer cells. We selected substrates from a phage display library of high diversity using secreted media of three established prostate cancer cell lines (DU-145, LNCaP and PC-3) with variable degrees of invasive capability. Some of these selected peptide substrates were hydrolyzed by the secreted proteins of all three prostatic cancer cell lines, demonstrating similarities in their proteolytic activities. On the other hand, a few substrates were cancer cell specific, indicating differences in the phenotypes of protease expression in prostate cancer. This work reports for the first time the selection of substrates from a mix of proteases using phage display technology and opens a new avenue for the direct identification of proteolytic activities for tumor extracts.
authors.affiliation: Urology Research Unit, Department of Urology, CHUV, Epalinges, Switzerland.
authors.forename: Sylvain M
authors.initials: SM
authors.lastname: Cloutier
authors.forename: Christoph
authors.initials: C
authors.lastname: Kündig
authors.forename: Christian M
authors.initials: CM
authors.lastname: Gygi
authors.forename: Patrice
authors.initials: P
authors.lastname: Jichlinski
authors.forename: Hans-Jürg
authors.initials: HJ
authors.lastname: Leisinger
authors.forename: David
authors.initials: D
authors.lastname: Deperthes
mesh_terms: D010450:Endopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D006868:Hydrolysis; D008297:Male; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000503:physiopathology*; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000494:pharmacology*; D019151:Peptide Library; D011471:Prostatic Neoplasms / Q000201:enzymology*; D011506:Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D019151:Peptide Library; D011506:Proteins; D010450:Endopeptidases; D010447:Peptide Hydrolases