Peptide from the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (TIMP-2) inhibits membrane activation of matrix metalloproteinase-2 (MMP-2)

Xiaoping Xu, Margarita Mikhailova, Zhihua Chen, Sanjay Pal, Trista K. Robichaud, Eileen M. Lafer, Sam Baber, Bjorn Steffensen

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Cellular activation of latent matrix metalloproteinase-2 (proMMP-2) requires formation of a cell membrane-associated activation complex that involves specific binding between the hemopexin domain of proMMP-2 (PEX) and the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (C-TIMP-2). In this study, we tested the feasibility of inhibiting activation of proMMP-2 by exogenous inhibitors, which block the binding between PEX and TIMP-2. The recombinant C-TIMP-2 and synthetic peptides from C-TIMP-2 were used as inhibitors for proMMP-2 activation. Recombinant C-TIMP-2 bound specifically to both the catalytically inactive MMP-2 E404A and the C-terminal domain of MMP-2 (PEX) in a concentration dependent manner with apparent K d of 3.9×10 -7M and 1.7×10 -7M, respectively. Moreover, C-TIMP-2 competed the binding between MMP-2 E404A and full-length TIMP-2. Finally, activity assays showed that addition of C-TIMP-2 to HT-1080 fibrosarcoma cells inhibited proMMP-2 activation in a concentration-dependent manner. We then designed a synthetic peptide, P175L, consisting of 20 residues from the PEX-binding tail region of C-TIMP-2. P175L bound PEX and inhibited cell membrane-mediated activation of proMMP-2 in a concentration dependent manner. Deletion of the last 9 tail residues of C-TIMP-2 in P175L abrogated the inhibitory activities of the peptide showing that these residues were essential for function. Overall, these experiments have demonstrated that proMMP-2 activation can be inhibited by exogenous inhibitors which points to a potential strategy for MMP-2 specific inhibition.

Original languageEnglish (US)
Pages (from-to)404-412
Number of pages9
JournalMatrix Biology
Volume30
Issue number7-8
DOIs
StatePublished - Sep 2011

Keywords

  • MMP inhibitor
  • MMP-2
  • Peptide
  • ProMMP-2 activation
  • Protein binding
  • TIMP-2

ASJC Scopus subject areas

  • Molecular Biology

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