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 alpha l and alpha 2 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, M., Monaco, S., Van Den Steen, P., Sbardella, D., Grasso, G., Marini, S., et al. (2009). The Collagen Binding Domain of Gelatinase A Modulates Degradation of Collagen IV by Gelatinase B. JOURNAL OF MOLECULAR BIOLOGY, 386(2), 419-434 [10.1016/j.jmb.2008.12.021].

The Collagen Binding Domain of Gelatinase A Modulates Degradation of Collagen IV by Gelatinase B

GIOIA, MAGDA;MARINI, STEFANO;COLETTA, MASSIMILIANO
2009-02-01

Abstract

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 alpha l and alpha 2 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
feb-2009
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/10 - BIOCHIMICA
English
Con Impact Factor ISI
collagen; collagen type 4; fibronectin; gelatinase A; gelatinase B; matrix metalloproteinase inhibitor; analytic method; article; binding site; catalysis; cell migration; collagen degradation; conformational transition; controlled study; edman sequencing; enzyme activity; human; human cell; mass spectrometry; neutrophil; placenta; priority journal; protein binding; protein degradation; protein domain; surface plasmon resonance; Collagen Type IV; Environmental Monitoring; Humans; Kinetics; Mass Spectrometry; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Peptides; Protein Binding; Protein Interaction Mapping; Sequence Analysis, Protein; Surface Plasmon Resonance; Temperature
Gioia, M., Monaco, S., Van Den Steen, P., Sbardella, D., Grasso, G., Marini, S., et al. (2009). The Collagen Binding Domain of Gelatinase A Modulates Degradation of Collagen IV by Gelatinase B. JOURNAL OF MOLECULAR BIOLOGY, 386(2), 419-434 [10.1016/j.jmb.2008.12.021].
Gioia, M; Monaco, S; Van Den Steen, P; Sbardella, D; Grasso, G; Marini, S; Overall, C; Opdenakker, G; Coletta, M
Articolo su rivista
File in questo prodotto:
File Dimensione Formato  
JMB2009CollIVCBD.pdf

accesso aperto

Dimensione 881.6 kB
Formato Adobe PDF
881.6 kB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/41241
Citazioni
  • ???jsp.display-item.citation.pmc??? 22
  • Scopus 46
  • ???jsp.display-item.citation.isi??? 45
social impact