Characterization of the mechanisms by which gelatinase A, neutrophil collagenase, and membrane-type metalloproteinase MMP-14 recognize collagen I and enzymatically process the two alpha-chains.
The turnover of native collagen has been ascribed to different members of the matrix metalloproteinase (MMP) family. Here, the mechanisms by which neutrophil collagenase (MMP-8), gelatinase A (MMP-2), and the ectodomain of MT1-MMP (ectMMP-14) degrade fibrillar collagen were examined. In particular, the hydrolysis of type I collagen at 37 degrees C was investigated to identify functional differences in the processing of the two alpha-chain types of fibrillar collagen. Thermodynamic and kinetic parameters were used for a quantitative comparison of the binding, unwinding, and hydrolysis of triple helical collagen. We demonstrate that the MMP family has developed at least two distinct mechanisms for collagen unwinding and cleavage. MMP-8 and ectMMP-14 display a similar mechanism (although with different catalytic parameters), which is characterized by binding (likely through the hemopexin-like domain) and cleavage of alpha-1 and/or alpha-2 chains without distinguishing between them and keeping the gross conformation of the triple helix (at least during the first cleavage step). On the other hand, MMP-2 binds preferentially the alpha-1 chains (likely through the fibronectin-like domain, which is not present in MMP-8 and ectMMP-14), grossly altering the whole triple helical arrangement of the collagen molecule and cleaving preferentially the alpha-2 chain. These distinctive mechanisms underly a drastically different mode of interaction with triple helical fibrillar collagen I, according to which the MMP domain is involved in binding. These findings can be related to the different role exerted by these MMPs on collagen homeostasis in the extracellular matrix.
Gioia
Magda
M
Department of Experimental Medicine and Biochemical Sciences, University of Roma Tor Vergata, Via Montpellier 1, I-00133 Rome, Italy.
Monaco
Susanna
S
Fasciglione
Giovanni Francesco
GF
Coletti
Anna
A
Modesti
Andrea
A
Marini
Stefano
S
Coletta
Massimo
M
D000818:Animals; D002384:Catalysis; D002417:Cattle; D024042:Collagen Type I / Q000737:chemistry*; D007700:Kinetics; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry*; D020778:Matrix Metalloproteinase 2 / Q000737:chemistry*; D020784:Matrix Metalloproteinase 8 / Q000737:chemistry*; D051379:Mice; D011487:Protein Conformation; D013816:Thermodynamics
D024042:Collagen Type I; D020778:Matrix Metalloproteinase 2; D020784:Matrix Metalloproteinase 8; D053511:Matrix Metalloproteinase 14