Matrix metalloproteinases process the laminin-5 γ2-chain and regulate epithelial cell migration

https://doi.org/10.1016/S0006-291X(03)00452-2Get rights and content

Abstract

Matrix metalloproteinase (MMP)-2 and membrane type 1-MMP can process the laminin-5 (Ln-5) γ2-chain, revealing a cryptic site inducing epithelial cell migration. We investigated whether other MMPs process the Ln-5 γ2-chain and related their ability to induce epithelial cell migration. The N-terminal sequences of the MMP-3, -12, -13, and -20 processed 80 kDa Ln-5 γ2x-chains were identical whereas the N-terminus of the 80kDaMMP-8 Ln-5 γ2x-chain was not. MMP-3, -13, -14, and -20 induced MCF-7 cell migration over Ln-5 while MMP-8 was a poor inducer of MCF-7 cell migration. In conclusion, several MMPs can process the Ln-5 γ2-chain and induce 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)

  • G. Murphy et al.

    Curr. Opin. Cell. Biol.

    (1999)
  • W.G. Carter et al.

    Cell

    (1991)
  • P. Rousselle et al.

    J. Biol. Chem.

    (1995)
  • S. Salo et al.

    Matrix Biol.

    (1999)
  • M. Makela et al.

    Exp. Cell. Res.

    (1999)
  • E. Pirila et al.

    Biochem. Biophys. Res. Commun.

    (2001)
  • Y.T. Konttinen et al.

    Matrix

    (1991)
  • H.S. Rasmussen et al.

    Pharmacol. Ther.

    (1997)
  • R. Hanemaaijer et al.

    J. Biol. Chem.

    (1997)
  • S. Amano et al.

    J. Biol. Chem.

    (2000)
  • H. Colognato et al.

    Dev. Dyn.

    (2000)
  • S.E. Baker et al.

    J. Cell. Sci.

    (1996)
  • G. Giannelli et al.

    Science

    (1997)
  • N. Koshikawa et al.

    J. Cell. Biol.

    (2000)
  • T. Kainulainen et al.

    J. Histochem. Cytochem.

    (1998)
  • Cited by (87)

    • Next generation matrix metalloproteinase inhibitors — Novel strategies bring new prospects

      2017, Biochimica et Biophysica Acta - Molecular Cell Research
      Citation Excerpt :

      Today, the matricryptins originating from collagen types I, IV, XVII, and XVIII are known [44–47]. Exposure of cryptic epitope during the γ2 chain of laminin-5 (Ln-5) degradation by MMP-2 -3, -8, -12, -13, -20 and MT1-MMP was shown to induce epithelial cell migration in vitro [48,49]. Since Ln-5 is widely spread in epithelial basement membranes, its cleavage by MMPs may be the general mechanism that triggers migration of cells in contact with epithelial basement membranes [48].

    • Towards integrating extracellular matrix and immunological pathways

      2017, Cytokine
      Citation Excerpt :

      In addition to elastin- and collagen- derived fragments, matrikines generated from cleavage of several other ECM proteoglycans and glycoproteins have been identified. Laminin, a major component of ECM basement membranes, can be cleaved by numerous MMPs and ADAM9 to generate matrikines that influence epithelial cell migration [97–99], neutrophil infiltration [100], and alveolar regeneration by engaging EGF receptors expressed on epithelial progenitor cells [101]. Proteolysis of the associated proteoglycans hyaluronan and versican also produces biologically active fragments that modulate inflammatory responses to infection and injury [102,103].

    View all citing articles on Scopus

    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.

    1

    These authors contributed equally to the manuscript.

    View full text