Cleavage at the stem region releases an active ectodomain of the membrane type 1 matrix metalloproteinase

Marta Toth, Pamela Osenkowski, Dusan Hesek, Stephen Brown, Samy Meroueh, Wael Sakr, Shahriar Mobashery, Rafael Fridman

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

MT1-MMP (membrane type 1 matrix metalloproteinase) is a membrane-anchored MMP that can be shed to the extracellular milieu. In the present study we report the primary structure and activity of the major soluble form of MT1-MMP. MS analysis of the purified 50-kDa soluble MT1-MMP form shows that the enzyme extends from Tyr112 to Val524, indicating that formation of this species requires a proteolytic cleavage within the stem region. In agreement, deletion of the entire stem region of MT1-MMP inhibited shedding of the 50-kDa species. A recombinant 50-kDa species (Tyr112-Val 524) expressed in cells exhibited enzymatic activity against pro-MMP-2 and galectin-3, and thus this species is a competent protease. The recombinant 50-kDa soluble form also decreased the level of surface-associated TIMP-2 (tissue inhibitor of metalloproteinase 2) when administered to cells expressing wild-type membrane-anchored MT1-MMP, suggesting that ectodomain shedding of MT1-MMP can alter the MMP/TIMP balance on the cell surface. A ∼53-kDa species of MT1-MMP was also isolated from a non-detergent extract of human breast carcinoma tissue and was found to lack the cytosolic tail, as determined with specific MT1-MMP domain antibodies. Together, these data show that MT1-MMP ectodomain shedding is a physiological process that may broaden MT1-MMP activity to the pericellular space.

Original languageEnglish (US)
Pages (from-to)497-506
Number of pages10
JournalBiochemical Journal
Volume387
Issue number2
DOIs
StatePublished - Apr 15 2005

Keywords

  • Ectodomain
  • Matrix metalloproteinase (MMP)
  • Protease
  • Proteolysis
  • Shedding
  • Tissue inhibitor of metalloproteinase (TIMP)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Toth, M., Osenkowski, P., Hesek, D., Brown, S., Meroueh, S., Sakr, W., Mobashery, S., & Fridman, R. (2005). Cleavage at the stem region releases an active ectodomain of the membrane type 1 matrix metalloproteinase. Biochemical Journal, 387(2), 497-506. https://doi.org/10.1042/BJ20041324