Journal
JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY
Volume 17, Issue 4, Pages 663-672Publisher
SPRINGER
DOI: 10.1007/s00775-012-0886-z
Keywords
Fibroblast collagenase; Collagen I; Collagenolytic cleavage mechanism; Kinetics; Hinge region
Funding
- Italian Space Agency [ASI 2005 OSMA]
- National Institutes of Health [AR-41843, CA-107183]
Ask authors/readers for more resources
The role of the hinge region in the unwinding and cleavage of type I collagen by interstitial collagenase (MMP-1) has been studied at 37 A degrees C and pH 7.3. The collagenolytic processing by MMP-1 displays a very similar overall rate for both chains of collagen I, even though the affinity is higher for the alpha-1 chain and the cleavage rate is faster for the alpha-2 chain. MMP-1 binding to collagen I brings about a significant unwinding of the triple-helical arrangement only after the first cleavage step of the alpha-1 and alpha-2 chains. The proteolytic processing by wild-type MMP-1 on a synthetic substrate and collagen I has been compared with that observed for site-directed mutants obtained either by truncating the hinge region (a dagger 255-272) or by individually replacing the conserved amino acids Val268, Gly272, and Lys277 of the hinge region with residues observed for the corresponding position in stromelysin-1 (MMP-3), a noncollagenolytic metalloproteinase. The a dagger 256-272 mutant has no collagenolytic activity, clearly demonstrating the crucial role of this region for the enzymatic processing of collagen I. However, among various mutants investigated, only Gly272Asp shows a dramatically reduced enzymatic activity both on the synthetic substrate and on collagen I. This effect, however, is clearly related to the substituting residue, since substitution of Ala or Asn for Gly272 does not have any effect on the kinetic properties of MMP-1. These data suggest that the substrate specificity of MMP-1 is dictated by the reciprocal structural relationships between the catalytic domain and the carboxy-terminal domain through the conformational arrangement of the hinge region.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available