[Show abstract][Hide abstract] ABSTRACT: The extracellular matrix macromolecule laminin-5 (Ln-5) is converted by matrix metalloproteinases (MMP) MT1-MMP and MMP-2 into a migration-promoting substrate in vitro. We now report that cleavage of Ln-5 by MT1-MMP occurs in vivo and affects epithelial tissue organization and probably Ln-5 turnover. In MT1-MMP knockout (KO) mice, the kidneys showed increased levels of total Ln-5 gamma2 subunit, but significantly reduced amounts of gamma2', an amino-terminal truncated proteolytic form of gamma2. The kidney tubular epithelia of KO animals were poorly differentiated, a phenotype reminiscent of human congenital mixed hypoplastic/dysplastic disorders. To establish a better link between Ln-5 proteolytic cleavage and epithelial morphology, MT1-MMP expression was reconstituted by transfection of MT1-MMP into a Ln-5 positive, MT1-MMP deficient epithelial cell line. MT1-MMP transfectants demonstrated increased levels of processed Ln-5 gamma2 chain and enhanced spreading on Ln-5, but not fibronectin. Recombinant MT1-MMP cleaved gamma2 constructs in vitro at a known in vivo gamma2 gamma2' processing site. These results strongly indicate that Ln-5 is a physiological substrate of MT1-MMP in vivo. Proteolytic processing of gamma2 subunit by MT1-MMP may influence Ln-5 turnover in epithelial basement membranes and affect epithelial morphogenesis.
[Show abstract][Hide abstract] ABSTRACT: Proteolytic cleavage of extracellular matrix (ECM) proteins by matrix metalloproteinases and/or conformational changes unmask "cryptic" sites and liberate fragments with biological activities that are not observed in the intact molecule. Cryptic sites and fragments of ECM macromolecules have been implicated in many events governed by cell-ECM interactions, such as migration, invasion, adhesion and differentiation. The unmasking of cryptic sites is a tightly controlled process, reflecting the importance of cryptic ECM functions. This review summarizes and evaluates the current developments regarding cryptic regulatory ECM signals found as ECM-tethered protein epitopes or fragments.
No preview · Article · Aug 2003 · Trends in Cell Biology
[Show abstract][Hide abstract] ABSTRACT: Extracellular matrix (ECM) fragments or cryptic sites unmasked by proteinases have been postulated to affect tissue remodeling and cancer progression. Therefore, the elucidation of their identities and functions is of great interest. Here, we show that matrix metalloproteinases (MMPs) generate a domain (DIII) from the ECM macromolecule laminin-5. Binding of a recombinant DIII fragment to epidermal growth factor receptor stimulates downstream signaling (mitogen-activated protein kinase), MMP-2 gene expression, and cell migration. Appearance of this cryptic ECM ligand in remodeling mammary gland coincides with MMP-mediated involution in wild-type mice, but not in tissue inhibitor of metalloproteinase 3 (TIMP-3)-deficient mice, supporting physiological regulation of DIII liberation. These findings indicate that ECM cues may operate via direct stimulation of receptor tyrosine kinases in tissue remodeling, and possibly cancer invasion.
Preview · Article · May 2003 · The Journal of Cell Biology
[Show abstract][Hide abstract] ABSTRACT: Laminins are a family of extracellular matrix glycoproteins involved in cell adhesion and migration. A major obstacle to understanding their structure-function relationships is the lack of small laminin domains capable of replicating integrin-binding, cell-adhesive, and migratory functions of the intact molecule. Here, we show that the recombinant LG3 (rLG3) module (26 kDa) of laminin-5 (Ln-5) alpha(3) chain replicated key Ln-5 activities. rLG3 but not rLG1 or rLG2 supported cell adhesion and migration of at least two distinct cell lines, in an integrin alpha(3)beta(1)-dependent manner. Cell adhesion to rLG3 was regulated by divalent cations and accompanied by cell spreading and tyrosine phosphorylation of FAK focal adhesion kinase. The integrin binding activity of rLG3 was confirmed by rLG3 affinity chromatography of detergent cell lysates, which resulted in specific purification of integrin alpha(3)beta(1). To our knowledge, this is the first report directly demonstrating that a recombinant laminin LG module is an active domain capable of supporting integrin-dependent cell adhesion and migration.
No preview · Article · Sep 2001 · Journal of Biological Chemistry