Mutational and structural analyses of the hinge region of membrane type 1-matrix metalloproteinase and enzyme processing

Pamela Osenkowski, Samy Meroueh, Dumitru Pavel, Shahriar Mobashery, Rafael Fridman

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Membrane type 1 (MT1)-matrix metalloproteinase (MMP) is a major mediator of collagen degradation in the pericellular space in both physiological and pathological conditions. Previous evidence has shown that on the cell surface, active MT1-MMP undergoes autocatalytic processing to a major membrane-tethered 44-kDa product lacking the catalytic domain and displaying Gly285 at its N terminus, which is at the beginning of the hinge domain. However, the importance of this site and the hinge region in MT1-MMP processing is unknown. In the current study, we generated mutations and deletions in the hinge of MT1-MMP and followed their effect on processing. These studies established Gly284-Gly285 as the main cleavage site involved in the formation of the 44-kDa species. However, alterations at this site did not prevent processing. Instead, they forced downstream cleavages within the stretch of residues flanked by Gln296 and Ser304 in the hinge region, as determined by the processing profile of various hinge deletion mutants. Also, replacement of the hinge of MT1-MMP with the longer MT3-MMP hinge did not prevent processing of MT1-MMP. Molecular dynamic studies using a computational model of MT1-MMP revealed that the hinge region is a highly motile element that undergoes significant motion in the highly exposed loop formed by Pro295-Arg302 consistent with being a prime target for proteolysis, in agreement with the mutational data. These studies suggest that the hinge of MT1-MMP evolved to facilitate processing, a promiscuous but compulsory event in the destiny of MT1-MMP, which may play a key role in the control of pericellular proteolysis.

Original languageEnglish (US)
Pages (from-to)26160-26168
Number of pages9
JournalJournal of Biological Chemistry
Volume280
Issue number28
DOIs
StatePublished - Jul 15 2005
Externally publishedYes

Fingerprint

Matrix Metalloproteinase 14
Hinges
Enzymes
Processing
Proteolysis
Matrix Metalloproteinase 16
Sequence Deletion
Molecular Dynamics Simulation
Molecular dynamics
Catalytic Domain
Collagen
Membranes
Degradation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mutational and structural analyses of the hinge region of membrane type 1-matrix metalloproteinase and enzyme processing. / Osenkowski, Pamela; Meroueh, Samy; Pavel, Dumitru; Mobashery, Shahriar; Fridman, Rafael.

In: Journal of Biological Chemistry, Vol. 280, No. 28, 15.07.2005, p. 26160-26168.

Research output: Contribution to journalArticle

Osenkowski, Pamela ; Meroueh, Samy ; Pavel, Dumitru ; Mobashery, Shahriar ; Fridman, Rafael. / Mutational and structural analyses of the hinge region of membrane type 1-matrix metalloproteinase and enzyme processing. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 28. pp. 26160-26168.
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