Enzymatic processing of collagen IV by MMP-2 (gelatinase A) affects neutrophil migration and it is modulated by extracatalytic domains

Protein Sci. 2006 Dec;15(12):2805-15. doi: 10.1110/ps.062430706. Epub 2006 Nov 6.


Proteolytic degradation of basement membrane influences the cell behavior during important processes, such as inflammations, tumorigenesis, angiogenesis, and allergic diseases. In this study, we have investigated the action of gelatinase A (MMP-2) on collagen IV, the major constituent of the basement membrane. We have compared quantitatively its action on the soluble forms of collagen IV extracted with or without pepsin (from human placenta and from Engelbreth-Holm-Swarm [EHS] murine sarcoma, respectively). The catalytic efficiency of MMP-2 is dramatically reduced in the case of the EHS murine sarcoma with respect to the human placenta, probably due to the much tighter packing of the network which renders very slow the speed of the rate-limiting step. We have also enquired on the role of MMP-2 domains in processing collagen IV. Addition of the isolated collagen binding domain, corresponding to the fibronectin-like domain of whole MMP-2, greatly in hibits the cleavage process, demonstrating that MMP-2 interacts with collagen type IV preferentially through its fibronectin-like domain. Conversely, the removal of the hemopexin-like domain, using only the catalytic domain of MMP-2, has only a limited effect on the catalytic efficiency toward collagen IV, indicating that the missing domain does not have great relevance for the overall mechanism. Finally, we have investigated the effect of MMP-2 proteolytic activity ex vivo. MMP-2 action negatively affects the neutrophils' migration across type IV coated membranes and this is likely related to the production of lower molecular weight fragments that impair the cellular migration.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Catalysis
  • Catalytic Domain / physiology*
  • Cell Membrane / metabolism
  • Cell Movement
  • Collagen Type IV / chemistry
  • Collagen Type IV / metabolism*
  • Female
  • Humans
  • Matrix Metalloproteinase 2 / chemistry*
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinase 2 / physiology
  • Mice
  • Mutant Proteins / chemistry
  • Mutant Proteins / metabolism
  • Neutrophils / cytology*
  • Placenta / enzymology
  • Placenta / metabolism
  • Protein Denaturation
  • Protein Structure, Tertiary / physiology
  • Sarcoma, Experimental / enzymology
  • Sarcoma, Experimental / metabolism


  • Collagen Type IV
  • Mutant Proteins
  • Matrix Metalloproteinase 2