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rdf:type |
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lifeskim:mentions |
umls-concept:C0014442,
umls-concept:C0017262,
umls-concept:C0086418,
umls-concept:C0145946,
umls-concept:C0172537,
umls-concept:C0185117,
umls-concept:C0439855,
umls-concept:C1880022,
umls-concept:C1880497,
umls-concept:C1996904,
umls-concept:C2911684
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pubmed:dateCreated |
1993-2-23
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pubmed:abstractText |
The human 72 kDa gelatinase/type IV collagenase is a metalloproteinase that is thought to play a role in metastasis and angiogenesis. The 72 kDa progelatinase can be isolated from conditioned media as a complex with the tissue inhibitor of metalloproteinase-2 (TIMP-2). To investigate 72 kDa gelatinase-TIMP-2 interactions and to compare the activity of the complex versus that of the free enzyme, we have expressed and purified human 72 kDa progelatinase and TIMP-2 as single proteins in a recombinant vaccinia virus mammalian cell expression system. The recombinant 72 kDa progelatinase was able to bind TIMP-2, and it digested gelatin and collagen type IV after activation by p-aminophenylmercuric acid (APMA). The specific activity of the recombinant free enzyme was 20-fold higher than the activity of an APMA-treated stoichiometric complex of recombinant 72 kDa progelatinase and TIMP-2. Also, TIMP-2 caused an 86% inhibition of activity when added to the activated enzyme at a 1:1 molar ratio. Activation of the free recombinant 72 kDa progelatinase yielded the 62 kDa species and two fragments of 46 and 35 kDa that cross-reacted with monoclonal antibodies to the 72 kDa proenzyme. TIMP-2 inhibited the conversion of the recombinant proenzyme to the 62 kDa species and the appearance of the 45 and 35 kDa bands. These results suggest that TIMP-2 is not only a potent inhibitor of the activated enzyme but also prevents the generation of low-molecular-mass species and full enzymic activity from the zymogen.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1310615,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1322396,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1645772,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1646720,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1699524,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1703045,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1849044,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1850424,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-1931943,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2009533,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2132731,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2169338,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2173706,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2174435,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2269296,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2380196,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2383557,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2461565,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2465572,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2536363,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2545347,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2548200,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2554304,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-271968,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2793861,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-2834383,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-3032947,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-3095828,
http://linkedlifedata.com/resource/pubmed/commentcorrection/8380993-3112559
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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 |
Jan
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pubmed:issn |
0264-6021
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
289 ( Pt 2)
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
411-6
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:8380993-Blotting, Western,
pubmed-meshheading:8380993-Chromatography, Affinity,
pubmed-meshheading:8380993-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8380993-Gelatinases,
pubmed-meshheading:8380993-HeLa Cells,
pubmed-meshheading:8380993-Humans,
pubmed-meshheading:8380993-Kinetics,
pubmed-meshheading:8380993-Molecular Weight,
pubmed-meshheading:8380993-Neoplasm Proteins,
pubmed-meshheading:8380993-Pepsin A,
pubmed-meshheading:8380993-Protein Binding,
pubmed-meshheading:8380993-Recombinant Proteins,
pubmed-meshheading:8380993-Tissue Inhibitor of Metalloproteinase-2
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pubmed:year |
1993
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pubmed:articleTitle |
Expression of human recombinant 72 kDa gelatinase and tissue inhibitor of metalloproteinase-2 (TIMP-2): characterization of complex and free enzyme.
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pubmed:affiliation |
Molecular Oncology Inc., Gaithersburg, MD 20878.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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