Source:http://linkedlifedata.com/resource/pubmed/id/15161915
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
31
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| pubmed:dateCreated |
2004-7-26
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| pubmed:abstractText |
Overexpression of hepatocyte growth factor (HGF) and its receptor Met often occurs in carcinoma cells, leading to establishment of an HGF/Met autocrine loop. Therefore, disruption of the HGF/Met autocrine loop may lead to down-regulation of tumorigenesis. To study the HGF/Met interaction, we have developed a cell-free system to detect HGF binding to a Met fusion protein, Met-IgG, using a modified enzyme-linked immunosorbent assay methodology. Since we previously showed that HGF can be purified by copper(II) affinity chromatography, we further explored the effect of copper(II) on the HGF/Met interaction. The divalent metal cations copper(II) and zinc(II) significantly inhibited HGF binding to immobilized Met-IgG with IC(50) values of 230-270 microM, respectively, whereas manganese(II) and magnesium(II) were less inhibitory with 20-60-fold higher IC(50) values. Incubation of 1 mM copper(II) with HGF resulted in nondenaturing and denaturing gel-mobility shifts, indicating that copper(II) binds directly to HGF. This interaction occurs at the N terminus of HGF, as incubation of 1 mM copper(II) with both HGF and the HGF derivative NK1 yielded similar results on SDS-PAGE. HGF-induced activation of Met and cell scattering were inhibited upon addition of HGF in the presence of 1 mM and 500 microM copper(II), respectively. Chemical protonation with diethyl pyrocarbonate of HGF histidine residues impeded the ability of 500 microM copper(II) to inhibit the binding of HGF to immobilized Met-IgG. Based on the NK1 domain structure, we propose that copper(II) may interact with HGF via the histidine residues in either N-terminal or kringle domains. The inhibition of HGF/Met interaction and subsequent downstream cellular functions may be through direct interference by copper(II), such as a change in charge or an induced local conformational change. This putative copper(II) binding domain may be the basis for developing potential inhibitors of HGF/Met binding and downstream functions and could lead to novel strategies for anti-cancer treatment.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Cations,
http://linkedlifedata.com/resource/pubmed/chemical/Copper,
http://linkedlifedata.com/resource/pubmed/chemical/Edetic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Hepatocyte Growth Factor,
http://linkedlifedata.com/resource/pubmed/chemical/Histidine,
http://linkedlifedata.com/resource/pubmed/chemical/Immunoglobulin G,
http://linkedlifedata.com/resource/pubmed/chemical/MET protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Magnesium,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Proto-Oncogene Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Proto-Oncogene Proteins c-met,
http://linkedlifedata.com/resource/pubmed/chemical/Protons,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Growth Factor,
http://linkedlifedata.com/resource/pubmed/chemical/Tyrosine,
http://linkedlifedata.com/resource/pubmed/chemical/Zinc
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| pubmed:status |
MEDLINE
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| pubmed:month |
Jul
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| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
30
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| pubmed:volume |
279
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
32499-506
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| pubmed:dateRevised |
2010-11-18
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| pubmed:meshHeading |
pubmed-meshheading:15161915-Animals,
pubmed-meshheading:15161915-Binding Sites,
pubmed-meshheading:15161915-Cations,
pubmed-meshheading:15161915-Cell Line,
pubmed-meshheading:15161915-Cell-Free System,
pubmed-meshheading:15161915-Copper,
pubmed-meshheading:15161915-Dogs,
pubmed-meshheading:15161915-Dose-Response Relationship, Drug,
pubmed-meshheading:15161915-Down-Regulation,
pubmed-meshheading:15161915-Edetic Acid,
pubmed-meshheading:15161915-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:15161915-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:15161915-Hepatocyte Growth Factor,
pubmed-meshheading:15161915-Histidine,
pubmed-meshheading:15161915-Immunoglobulin G,
pubmed-meshheading:15161915-Inhibitory Concentration 50,
pubmed-meshheading:15161915-Magnesium,
pubmed-meshheading:15161915-Models, Molecular,
pubmed-meshheading:15161915-Phosphorylation,
pubmed-meshheading:15161915-Protein Binding,
pubmed-meshheading:15161915-Protein Structure, Tertiary,
pubmed-meshheading:15161915-Proteins,
pubmed-meshheading:15161915-Proto-Oncogene Proteins,
pubmed-meshheading:15161915-Proto-Oncogene Proteins c-met,
pubmed-meshheading:15161915-Protons,
pubmed-meshheading:15161915-Receptors, Growth Factor,
pubmed-meshheading:15161915-Tyrosine,
pubmed-meshheading:15161915-Zinc
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| pubmed:year |
2004
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| pubmed:articleTitle |
Inhibition by copper(II) binding of hepatocyte growth factor (HGF) interaction with its receptor Met and blockade of HGF/Met function.
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| pubmed:affiliation |
Cancer Research Institute, Division of Cancer Biology and Genetics, Queen's University, Kingston, Ontario, Canada.
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| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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