Catalytic activities and substrate specificity of the human membrane type 4 matrix metalloproteinase catalytic domain

Yahong Wang, Adam R. Johnson, Qizhuang Ye, Richard D. Dyer

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Membrane type (MT)-matrix metalloproteinases (MMPs) are recently recognized members of the family of Zn2+- and Ca2+-dependent MMPs. To investigate the proteolytic capabilities of human MT4-MMP (i.e. MMP-17), we have cloned DNA encoding its catalytic domain (CD) from a breast carcinoma cDNA library. Human membrane type 4 MMP CD (MT4-MMPCD) protein, expressed as inclusion bodies in Escherichia coli, was purified to homogeneity and refolded in the presence of Zn2+ and Ca2+. While MT4-MMPCD cleaved synthetic MMP substrates Ac-PLG-[2-mercapto-4-methylpentanoyl]-LG-OEt and Mca-PLGL-Dpa-AR-NH2 with modest efficiency, it catalyzed with much higher efficiency the hydrolysis of a pro-tumor necrosis factor-α converting enzyme synthetic substrate, Mca-PLAQAV-Dpa-RSSSR-NH2. Catalytic efficiency with the pro-tumor necrosis factor-α converting enzyme substrate was maximal at pH 7.4 and was modulated by three ionizable enzyme groups (pK(α3) = 6.2, pK(α2) = 8.3, and pK(α1) = 10.6). MT4-MMPCD cleaved gelatin but was inactive toward type I collagen, type IV collagen, fibronectin, and laminin. Like all known MT-MMPs, MT4-MMPCD was also able to activate 72-kDa progelatinase A to its 68-kDa form. EDTA, 1,10-phenanthroline, reference hydroxamic acid MMP inhibitors, tissue inhibitor of metalloproteinases-1, and tissue inhibitor of metalloproteinases-2 all potently blocked MT4-MMPCD enzymatic activity. MT4-MMP is, therefore, a competent Zn2+-dependent MMP with unique specificity among synthetic substrates and the capability to both degrade gelatin and activate progelatinase A.

Original languageEnglish (US)
Pages (from-to)33043-33049
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number46
DOIs
StatePublished - Nov 12 1999
Externally publishedYes

Fingerprint

Matrix Metalloproteinase 17
Substrate Specificity
Matrix Metalloproteinases
Catalyst activity
Catalytic Domain
Substrates
Membrane-Associated Matrix Metalloproteinases
Membranes
Gelatin
Enzymes
Tumor Necrosis Factor-alpha
Hydroxamic Acids
Tissue Inhibitor of Metalloproteinase-2
Tissue Inhibitor of Metalloproteinase-1
Matrix Metalloproteinase Inhibitors
Collagen Type IV
Inclusion Bodies
Laminin
Gene Library
Fibronectins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Catalytic activities and substrate specificity of the human membrane type 4 matrix metalloproteinase catalytic domain. / Wang, Yahong; Johnson, Adam R.; Ye, Qizhuang; Dyer, Richard D.

In: Journal of Biological Chemistry, Vol. 274, No. 46, 12.11.1999, p. 33043-33049.

Research output: Contribution to journalArticle

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