Biochemical and Biophysical Research Communications
Matrix metalloproteinases process the laminin-5 2-chain and regulate epithelial cell migration☆
Section snippets
Materials and methods
Materials. The MMPs used in this study were purchased as follows: MT1-MMP and human MMP-13 (Invitek GmbH, Germany); MMP-3 and -8 (Sigma Chemicals, St. Louis, MO); human MMP-7 (Chemicon International, Temecula, CA); and MMP-12 (Elastin Products Company, Owensville, MO). Human MMP-2 was purified from human gingival fibroblasts [18], human MMP-8 from extracts of polymorphonuclear neutrophils [19], human proMMP-3 from human synovial fibroblasts [20], and human proMMP-13 was prepared as described
Cleavage of the Ln-5 γ2-chain by MMPs
Ln-5 γ2-chain was incubated with MMP-3, -7, -8, -12, -13, -14, and -20 at a 1:3 enzyme:substrate ratio and the time-dependent effects were analyzed by Western immunoblotting with a Ln-5 γ2-chain specific antibody (Figs. 1A–F). MMPs increased Ln-5 γ2-chain processing dose-dependently (not shown). Unprocessed Ln-5 γ2-chain was present in the 140 kDa and mature 100 kDa forms (Figs. 1A–G).
Within 1 h, enamelysin (MMP-20) processed the 140 and 100 kDa forms of Ln-5 γ2-chain into 80 and 66 kDa forms (Fig.
Discussion
The present findings evidence that in addition to the previously published MMP-2 and MT1-MMP, several other MMPs can also process the Ln-5 γ2-chain and reveal the cryptic site inducing increased transformed epithelial cell migration.
The 140-kDa form of the Ln-5 γ2-chain is proteolytically processed into a 100-kDa mature form [4]. Previous studies show that at least bone morphogenetic protein 1 and MT1-MMP can accomplish this cleavage [8], [30]. The N-terminal sequence of the 100 kDa Ln-5
Acknowledgements
This work was supported by grants from the Finnish Academy, the Finnish Dental Society Apollonia, the Else and Willhelm Stockman Foundation, the HUCH-EVO (TI020Y0002) grant, the Helsinki University Research Funds, the Biomedicum Helsinki Foundation, the K. Albin Johansson Foundation, Einar and Karin Stroems Foundation. This work was equally supervised by Timo Sorsa, Päivi Maisi, and Tuula Salo. Part of this work has been presented as an abstract (14, page 50) at the XVIII Federation of European
References (30)
- et al.
Curr. Opin. Cell. Biol.
(1999) - et al.
Cell
(1991) - et al.
J. Biol. Chem.
(1995) - et al.
Matrix Biol.
(1999) - et al.
Exp. Cell. Res.
(1999) - et al.
Biochem. Biophys. Res. Commun.
(2001) - et al.
Matrix
(1991) - et al.
Pharmacol. Ther.
(1997) - et al.
J. Biol. Chem.
(1997) - et al.
J. Biol. Chem.
(2000)
Dev. Dyn.
J. Cell. Sci.
Science
J. Cell. Biol.
J. Histochem. Cytochem.
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Abbreviations: MMP, matrix metalloproteinase; ECM, extracellular matrix; BM, basement membrane; APMA, p-aminophenylmercuric acetate; Ln-5, laminin-5; SCC, squamous cell carcinoma; MCF, mammary adenocarcinoma cells.
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These authors contributed equally to the manuscript.