Binding of gelatinases A and B to type-I collagen and other matrix components

Abstract

Matrix sequestration of matrix metalloproteinases may be important for the facilitation of remodelling events and the migration of cells through the extracellular matrix. Using an ELISA technique we studied the ability of pro and active forms of gelatinases A and B (GLA and GLB) to bind to matrix components and the contribution made by the different enzyme domains. Pro and active forms of GLA and GLB bound to type-I and type-IV collagens, gelatin and laminin films. Binding to collagens occurred exclusively via the N-terminal portion of the molecule in both of the gelatinases; deletion of the fibronectin-like domain in GLA abolished binding. Fibronectin was shown to compete with GLA, confirming that binding occurs through this domain. GLA and GLB competed for binding to collagen type I, whereas collagenase and stromelysin bound to different sites and could be co-localized with the gelatinases. We conclude that gelatinases have different binding specificities from those previously documented for stromelysin and collagenase, which bind through their C-terminal domains to collagen fibrils.

Type IV Collagen Induces Matrix Metalloproteinase 2 Activation in HT1080 Fibrosarcoma Cells

“…Indeed, in agreement with our observations (Fig. 3), Allan and co-workers [69] demonstrated that both pro-and active MMP-2 efficiently bind to thin coats of type I and type IV collagens, but very poorly to fibronectin. This collagen-binding property is solely mediated by the fi-bronectin-type II inserts present in the catalytic domain of the enzyme [69].…”

“…3), Allan and co-workers [69] demonstrated that both pro-and active MMP-2 efficiently bind to thin coats of type I and type IV collagens, but very poorly to fibronectin. This collagen-binding property is solely mediated by the fi-bronectin-type II inserts present in the catalytic domain of the enzyme [69]. As a consequence, the interaction of cell surface anchored MMP-2 with the molecules of type IV collagen is likely to induce the clustering of the MT1-MMP/TIMP-2/MMP-2 ternary complexes at the sites of interaction with the ECM, therefore promoting the intermolecular autocatalytic cleavage required to generate fully mature MMP-2.…”

Comparative analysis of the noncollagenous NC1 domain of type IV collagen: Identification of structural features important for assembly, function, and pathogenesis

“…It is also possible that the interaction of NCl with MMP-2 or MMP-3 might be weakened or abolished by the interaction of gelatin or transferrin substrates with the enzymes. Complex interactions of NCl have been reported with gelatin and other matrix components (Allan et al, 1995). Inhibition of both MMP-2 and MMP-3 in peptide assays argues against an involvement of the fibronectin-like domains which are unique to MMP-2.…”

The cell biology of leukocyte-mediated proteolysis

“…However, the susceptibility of these matrix-bound MMPs to inhibition were not examined in these studies. It is noteworthy in this respect that immunolocalization studies have demonstrated that MMP-2, MMP-9, MMP-3, and MMP-1 bind to matrix components in rapidly resorbing tissues in vivo [127], and it has been postulated that matrix binding of MMPs may confer retention, stability, and bioactivity for prolonged periods, thus facilitating their roles in pericellular proteolysis [125].…”

“…Both gelatinases (MMP-2 and MMP-9) have been shown to bind to type I and IV collagen, gelatin, and fibronectin via their amino-terminal domains [125]. Collagenase (MMP-1) and stromelysin (MMP-3) bind through their carboxy-terminal domains to collagen fibrils in vitro [126].…”