Pericellular activation of proMMP-7 (promatrilysin-1) through interaction with CD151.

Abstract

Matrix metalloproteinase-7 (MMP-7) (also known as matrilysin-1) is secreted as a proenzyme (proMMP-7) and plays a key role in the degradation of various extracellular matrix (ECM) and non-ECM molecules after activation. To identify the binding proteins related to proMMP-7 activation, a human lung cDNA library was screened by yeast two-hybrid system using proMMP-7 as bait. We identified a candidate molecule CD151, which is a member of the transmembrane 4 superfamily. Complex formation of proMMP-7 with CD151 was demonstrated by immunoprecipitation of the molecules from CaR-1 cells, a human rectal carcinoma cell line, expressing both proMMP-7 and CD151, and CD151 stable transfectants incubated with proMMP-7. Yeast two-hybrid assays using deletion mutants of proMMP-7 and CD151 suggested an interaction between the propeptide of proMMP-7 and the COOH-terminal extracellular loop of CD151. The binding activity of (125)I-labeled proMMP-7 to CD151 on the cell membranes was shown with CD151 stable transfectants. Laser-scanning confocal microscopy demonstrated that proMMP-7 colocalizes with CD151 on the cell membranes of CD151 stable transfectants and CaR-1 cells. In situ zymography using crosslinked carboxymethylated transferrin, a substrate of MMP-7, demonstrated proteinase activity on and around CD151 stable transfectants and CaR-1 cells, while the activity was abolished by their treatment with MMP inhibitors, anti-MMP-7 antibody or anti-CD151 antibody. In human lung adenocarcinoma tissues, colocalization of MMP-7 and CD151 was demonstrated on the carcinoma cells. Metalloproteinase activity was present in these tissues and could be inhibited by antibodies to MMP-7 or CD151. These data demonstrate for the first time that proMMP-7 is captured and activated on the cell membranes through interaction with CD151, and suggest the possibility that similar to the MT1-MMP/MMP-2 system, MMP-7 is involved in the pericellular activation mechanism mediated by CD151, a crucial step in proteolysis on the cell membranes under various pathophysiological conditions including cancer invasion and metastasis.

• Authors: Takayuki Shiomi, Isao Inoki, Fumio Kataoka, Takashi Ohtsuka, Gakuji Hashimoto, Ryoichi Nemori, Yasunori Okada
• Journal: Laboratory investigation; a journal of technical methods and pathology (Lab Invest) Vol. 85 Issue 12 Pg. 1489-506 (Dec 2005) ISSN: 0023-6837 [Print] United States
• PMID: 16200075 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Blocking
• Antigens, CD
• CD151 protein, human
• Enzyme Inhibitors
• Tetraspanin 24
• Matrix Metalloproteinase 7

Topics

• Adenocarcinoma (enzymology, pathology)
• Antibodies, Blocking (pharmacology)
• Antigens, CD (genetics, immunology, metabolism)
• Cell Line, Tumor
• Cell Membrane (drug effects, enzymology)
• Enzyme Activation
• Enzyme Inhibitors (pharmacology)
• Extracellular Matrix (metabolism)
• Gene Expression Regulation, Neoplastic
• Humans
• Lung Neoplasms (enzymology, pathology)
• Matrix Metalloproteinase 7 (genetics, immunology, metabolism)
• Neoplasm Invasiveness (physiopathology)
• Neoplasm Metastasis (physiopathology)
• Rectal Neoplasms (enzymology)
• Tetraspanin 24
• Transfection
• Two-Hybrid System Techniques