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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
28
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pubmed:dateCreated |
1998-8-12
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pubmed:abstractText |
We have undertaken a detailed analysis of the mechanism of inhibition of matrix metalloproteinase-2 (gelatinase A) by tissue inhibitor of metalloproteinase-2 (TIMP-2). Quenched fluorescent substrates have been used to analyze the rate of inhibition of gelatinase A by TIMP-2 over a wide range of TIMP-2 concentrations. When the values of the observed rate constant for the inhibition are plotted against TIMP-2 concentration, saturation is observed at high concentrations, providing evidence for formation of an intermediate in the pathway. Rate constants for the formation and dissociation of the intermediate are 5.9 x 10(6) M-1 s-1 and 6.3 s-1 respectively, giving a Ki for the initial step of approximately 1 microM. The rate constant for the association of the final complex is 33 s-1. By studying the dissociation of 125I-labeled TIMP-2 from a gelatinase A-TIMP-2 complex using ligand exchange experiments, we obtained a rate constant for the dissociation of the final stable complex of 2 x 10(-)8 s-1. This gives a value for the overall dissociation constant of approximately 0.6 fM.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0006-2960
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
14
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pubmed:volume |
37
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
10094-8
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:9665714-Binding, Competitive,
pubmed-meshheading:9665714-Enzyme Activation,
pubmed-meshheading:9665714-Fluorescent Dyes,
pubmed-meshheading:9665714-Gelatinases,
pubmed-meshheading:9665714-Humans,
pubmed-meshheading:9665714-Iodine Radioisotopes,
pubmed-meshheading:9665714-Kinetics,
pubmed-meshheading:9665714-Macromolecular Substances,
pubmed-meshheading:9665714-Matrix Metalloproteinase 2,
pubmed-meshheading:9665714-Metalloendopeptidases,
pubmed-meshheading:9665714-Oligopeptides,
pubmed-meshheading:9665714-Protein Binding,
pubmed-meshheading:9665714-Substrate Specificity,
pubmed-meshheading:9665714-Tissue Inhibitor of Metalloproteinase-2
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pubmed:year |
1998
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pubmed:articleTitle |
Kinetic analysis of the mechanism of interaction of full-length TIMP-2 and gelatinase A: evidence for the existence of a low-affinity intermediate.
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pubmed:affiliation |
School of Biological Sciences, University of East Anglia, Norwich, UK. m.hutton@uea.ac.uk
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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