The collagen binding domain of gelatinase A modulates degradation of collagen IV by gelatinase B.
Type IV collagen remodeling plays a critical role in inflammatory responses, angiogenesis and metastasis. Its remodeling is executed by a family of matrix metalloproteinases (MMPs), of which the constitutive gelatinase A (MMP2) and the inducible gelatinase B (MMP9) are key examples. Thus, in many pathological conditions, both gelatinases act together. Kinetic data are reported for the enzymatic processing at 37 degrees C of type IV collagen from human placenta by MMP9 and its modulation by the fibronectin-like collagen binding domain (CBD) of MMP2. The alpha1 and alpha2 chain components of type IV collagen were cleaved by gelatinases and identified by mass spectrometry as well as Edman sequencing. Surface plasmon resonance interaction assays showed that CBD bound type IV collagen at two topologically distinct sites. On the basis of linked-function analysis, we demonstrated that CBD of MMP2 tuned the cleavage of collagen IV by MMP9, presumably by inducing a ligand-linked structural change on the type IV collagen. At low concentrations, the CBD bound the first site and thereby allosterically modulated the binding of MMP9 to collagen IV, thus enhancing the collagenolytic activity of MMP9. At high concentrations, CBD binding to the second site interfered with MMP9 binding to collagen IV, acting as a competitive inhibitor. Interestingly, modulation of collagen IV degradation by inactive forms of MMP2 also occurred in a cell-based system, revealing that this interrelationship affected neutrophil migration across a collagen IV membrane. The regulation of the proteolytic processing by a catalytically inactive domain (i.e., CBD) suggests that the two gelatinases might cooperate in degrading substrates even when either one is inactive. This observation reinforces the idea of exosite targets for MMP inhibitors, which should include all macromolecular substrate recognition sites.
Gioia
Magda
M
Department of Experimental Medicine and Biochemical Sciences, University of Roma Tor Vergata, Rome, Italy.
Monaco
Susanna
S
Van Den Steen
Philippe E
PE
Sbardella
Diego
D
Grasso
Giuseppe
G
Marini
Stefano
S
Overall
Christopher M
CM
Opdenakker
Ghislain
G
Coletta
Massimo
M
D024141:Collagen Type IV / Q000378:metabolism*; D004784:Environmental Monitoring; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D010455:Peptides / Q000302:isolation & purification; D011485:Protein Binding; D025941:Protein Interaction Mapping; D020539:Sequence Analysis, Protein; D020349:Surface Plasmon Resonance; D013696:Temperature
D024141:Collagen Type IV; D010455:Peptides; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9