Abstract
A truncated form of the membrane-type matrix metalloproteinase-1 [(Ala21-Ile318)proMT1-MMP] lacking the hemopexin-like and trans-membrane domain was produced in E. coli. We demonstrate that the recombinant proenzyme was autoproteolytically processed to a fully active catalytic domain with N-terminal Ile114. The catalytic domain of MT1-MMP initiated the activation of progelatinase A and progelatinase A complexed with tissue inhibitor of metalloproteinases-2 (TIMP-2). As a typical soluble metalloproteinase it was able to cleave physiologic as well as synthetic substrates. Our kinetic data demonstrate that TIMP-2 is a potent inhibitor for the recombinant enzyme.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites
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Enzyme Activation
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Enzyme Precursors / metabolism*
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Gelatinases / metabolism*
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Kinetics
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Matrix Metalloproteinases, Membrane-Associated
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Metalloendopeptidases / chemistry
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Metalloendopeptidases / metabolism*
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Oligopeptides / metabolism
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Polymerase Chain Reaction
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Protease Inhibitors / metabolism
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Protease Inhibitors / pharmacology
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Proteins / metabolism*
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Proteins / pharmacology
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Tissue Inhibitor of Metalloproteinase-2
Substances
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Enzyme Precursors
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Oligopeptides
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Protease Inhibitors
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Proteins
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Recombinant Proteins
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Tissue Inhibitor of Metalloproteinase-2
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Gelatinases
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Matrix Metalloproteinases, Membrane-Associated
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Metalloendopeptidases
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progelatinase