TIMP-2 promotes activation of progelatinase A by membrane-type 1 matrix metalloproteinase immobilized on agarose beads

T. Kinoshita, H. Sato, [No Value] Akiko, [No Value] Okada, E. Ohuchi, K. Imai, Y. Okada, M. Seiki

Research output: Contribution to journalArticle

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Abstract

Membrane-type 1 matrix metalloproteinase (MT1-MMP)/MMP-14 is the activator of progelatinase A (proGelA)/proMMP-2 on the cell surface. However, it was a paradox that a tissue inhibitor of metalloproteinase-2 (TIMP-2), which is an inhibitor of MT1-MMP, is required for proGelA activation by the cells expressing MT1- MP. In this study, a truncated MT1-MMP having a FLAG- tag sequence at the C terminus (MT1-F) was immobilized onto agarose beads (MT1-F/B) and used to analyze the role of TIMP-2. The proteolytic activity of MT1-F/B against a synthetic peptide substrate was inhibited by TIMP-2 in a dose-dependent manner. In contrast, TIMP-2 promoted the processing of proGelA by MT1-F/B at low concentrations and inhibited it at higher concentrations. TIMP-2 promoted the binding of proGelA to the MT1-F on the beads by forming a trimolecular complex, which was followed by processing of proGelA. A stimulatory effect of TIMP-2 was observed under conditions in which unoccupied MT1-F was still available. Thus, the ternary complex is thought to act as a means to concentrate the substrate to the bead surface and to present it to the neighboring free MT1-F.

Original languageEnglish
Pages (from-to)16098-16103
Number of pages6
JournalJournal of Biological Chemistry
Volume273
Issue number26
DOIs
Publication statusPublished - 1998 Jun 26

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Matrix Metalloproteinase 14
Tissue Inhibitor of Metalloproteinase-2
Sepharose
Chemical activation
Substrates
Processing
Matrix Metalloproteinases
progelatinase
Cells
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

TIMP-2 promotes activation of progelatinase A by membrane-type 1 matrix metalloproteinase immobilized on agarose beads. / Kinoshita, T.; Sato, H.; Akiko, [No Value]; Okada, [No Value]; Ohuchi, E.; Imai, K.; Okada, Y.; Seiki, M.

In: Journal of Biological Chemistry, Vol. 273, No. 26, 26.06.1998, p. 16098-16103.

Research output: Contribution to journalArticle

Kinoshita, T, Sato, H, Akiko, NV, Okada, NV, Ohuchi, E, Imai, K, Okada, Y & Seiki, M 1998, 'TIMP-2 promotes activation of progelatinase A by membrane-type 1 matrix metalloproteinase immobilized on agarose beads', Journal of Biological Chemistry, vol. 273, no. 26, pp. 16098-16103. https://doi.org/10.1074/jbc.273.26.16098
Kinoshita, T. ; Sato, H. ; Akiko, [No Value] ; Okada, [No Value] ; Ohuchi, E. ; Imai, K. ; Okada, Y. ; Seiki, M. / TIMP-2 promotes activation of progelatinase A by membrane-type 1 matrix metalloproteinase immobilized on agarose beads. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 26. pp. 16098-16103.
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