Abstract
Peroxynitrite (ONOO-) has recently been implicated in connective tissue destruction in vivo. We have studied the effect of ONOO- on the activity of tissue inhibitor of metalloproteinase-1 (TIMP-1) in vitro. The inactivation of TIMP-1 by ONOO- was dose dependent with 50 microM ONOO- reducing the inhibitory activity of TIMP-1 towards gelatinase-A by 50%. High concentrations of ONOO- (500 microM-5 mM) caused protein fragmentation whilst lower concentrations (<250 microM) inactivated TIMP-1 without altering the molecular weight. Inactivation could be blocked by ONOO- scavengers but not by hydroxyl radical scavengers. Our results show that ONOO- is capable of inactivating TIMP-1, a process which could potentiate metalloproteinase-mediated tissue breakdown.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Amino Acids / pharmacology
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Electrophoresis, Polyacrylamide Gel
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Free Radical Scavengers / pharmacology
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Glycoproteins / antagonists & inhibitors*
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Glycoproteins / isolation & purification
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Glycoproteins / pharmacology*
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Humans
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Kinetics
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Matrix Metalloproteinase 3
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Metalloendopeptidases / antagonists & inhibitors
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Metalloendopeptidases / metabolism*
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Molecular Sequence Data
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Nitrates / chemical synthesis
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Nitrates / pharmacology*
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Recombinant Proteins / antagonists & inhibitors
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / pharmacology
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Substrate Specificity
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Tissue Inhibitor of Metalloproteinases
Substances
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Amino Acids
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Free Radical Scavengers
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Glycoproteins
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Nitrates
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Recombinant Proteins
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Tissue Inhibitor of Metalloproteinases
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peroxynitric acid
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Metalloendopeptidases
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Matrix Metalloproteinase 3