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Identification of substrates of the extracellular protease ADAMTS1 by DIGE proteomic analysis
Author(s) -
Canals Francesc,
Colomé Nuria,
Ferrer Cristina,
PlazaCalonge María del Carmen,
RodríguezManzaneque Juan Carlos
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.200500446
Subject(s) - proteases , thrombospondin , microbiology and biotechnology , biology , extracellular matrix , proteomics , metalloproteinase , extracellular , protease , glycoprotein , chemistry , matrix metalloproteinase , biochemistry , enzyme , gene
Proteolytic modification of components of the extracellular milieu by metalloproteinases plays important roles in the regulation of multiple cellular and physiological processes and pathological conditions. ADAMTS1 is a secreted enzyme of the ADAMTS ( a d isintegrin a nd m etalloproteinase with t hrombo s pondin motifs) family of proteases, which is related to angiogenesis and inflammation processes. Here, we describe a proteomic screening for putative ADAMTS1 substrates by analyzing the protein profiles obtained from cultures of transfected cells overexpressing the protease as compared to parental cells. Conditioned medium proteins of cultures of the two cell lines have been quantitatively compared by DIGE. Proteins showing differential levels have been identified by MS techniques leading to the finding of five potential new substrates of ADAMTS1: the basement membrane proteins nidogen‐1 and ‐2, the desmosomal protein desmocollin‐3, and the extracellular glycoproteins dystroglycan 1 and Mac‐2‐binding protein. Nidogen‐1 and ‐2 have been further validated as substrates by immunochemical analysis. Our results demonstrate the utility of the DIGE proteomic technique for the discovery of specific substrates of matrix proteases.