Abstract
We previously reported that endostatin inhibits endothelial and tumor cellular invasion by blocking activation and catalytic activity of matrix metalloproteinase (MMP)-2. Here we have examined the domain of proMMP-2 responsible for the binding of endostatin using surface plasmon resonance. ProMMP-2 and proMMP-2deltaHP lacking the hinge and hemopexin-like (HP) domains bound little to the immobilized endostatin. The active MMP-2 and MMP-2deltaHP, but not the HP domain of MMP-2, bound to endostatin at similar levels. In addition, preincubation of MMP-2 and MMP-2deltaHP with the MMP inhibitor actinonin, which binds to the active site of MMP-2, abolished their bindings to endostatin. These results indicate that endostatin binds to neither the latent proMMP-2 nor the HP domain but to the catalytic domain of MMP-2.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Catalytic Domain / physiology
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Cell Line
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Collagen / genetics
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Collagen / metabolism*
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Endostatins
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Enzyme Inhibitors / pharmacology
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Enzyme Precursors / antagonists & inhibitors
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Enzyme Precursors / genetics
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Enzyme Precursors / metabolism
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Gelatinases / antagonists & inhibitors
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Gelatinases / genetics
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Gelatinases / metabolism
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Humans
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Hydroxamic Acids / pharmacology
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Matrix Metalloproteinase 2 / metabolism*
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Matrix Metalloproteinase Inhibitors
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Metalloendopeptidases / antagonists & inhibitors
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Metalloendopeptidases / genetics
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Metalloendopeptidases / metabolism
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Mice
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Peptide Fragments / genetics
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Peptide Fragments / metabolism*
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Protein Binding / physiology
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Protein Structure, Tertiary / physiology
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Sequence Deletion
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Spodoptera
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Surface Plasmon Resonance
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Tissue Inhibitor of Metalloproteinase-2 / genetics
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Tissue Inhibitor of Metalloproteinase-2 / metabolism
Substances
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Endostatins
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Enzyme Inhibitors
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Enzyme Precursors
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Hydroxamic Acids
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Matrix Metalloproteinase Inhibitors
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Peptide Fragments
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Tissue Inhibitor of Metalloproteinase-2
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Collagen
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Gelatinases
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Metalloendopeptidases
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progelatinase
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Matrix Metalloproteinase 2
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actinonin