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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2001-7-2
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pubmed:abstractText |
Activated receptor tyrosine kinases induce a large number of tyrosine phosphorylation-dependent protein-protein interactions through which they mediate their various ligand-exerted functions including regulation of proliferation, differentiation and survival. TrkB receptor tyrosine kinase activated by binding of brain-derived neurotrophic factor (BDNF) also stimulates various protein interactions in a tyrosine phosphorylation-dependent manner in neuronal cells. To examine tyrosine phosphorylation-dependent interactions stimulated by active TrkB, we developed a modified yeast two-hybrid system, which we call the yeast two-and-a-half-hybrid system. In this system, yeast was engineered to express a tyrosine kinase domain of TrkB as an effector, in addition to two fusion proteins with GAL4 DNA-binding and GAL4 activation domains as bait and prey proteins, respectively. Using this system with Shp2 as the bait, we demonstrated that Shp2 interacts directly with BIT/SHPS-1 (also called SIRP) and Grb2 depending on tyrosine phosphorylation mediated by TrkB. Furthermore, we screened an adult human brain cDNA library with the yeast two-and-a-half-hybrid system in order to identify other Shp2-binding proteins in TrkB-stimulated tyrosine phosphorylation signaling. We found that fibroblast growth factor receptor substrate 2beta (FRS2beta), also called SNT2, interacts with Shp2 dependently on TrkB-mediated tyrosine phosphorylation of FRS2beta/SNT2. Therefore, we show that the two-and-a-half-hybrid system is a powerful tool for studying tyrosine phosphorylation-dependent protein-protein interactions in intracellular signaling pathways stimulated by TrkB receptor tyrosine kinase.
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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 |
http://linkedlifedata.com/resource/pubmed/chemical/Adaptor Proteins, Signal Transducing,
http://linkedlifedata.com/resource/pubmed/chemical/Antigens, Differentiation,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/FRS2 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/GRB2 Adaptor Protein,
http://linkedlifedata.com/resource/pubmed/chemical/GRB2 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Intracellular Signaling Peptides...,
http://linkedlifedata.com/resource/pubmed/chemical/Lipoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Glycoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Membrane Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Neural Cell Adhesion Molecule L1,
http://linkedlifedata.com/resource/pubmed/chemical/Neural Cell Adhesion Molecules,
http://linkedlifedata.com/resource/pubmed/chemical/PTPN11 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/PTPN6 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphoproteins,
http://linkedlifedata.com/resource/pubmed/chemical/Phosphotyrosine,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Tyrosine Phosphatase...,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Tyrosine Phosphatase...,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Tyrosine Phosphatases,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Receptor, trkB,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Immunologic
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0021-924X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
130
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
157-65
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:11432792-Adaptor Proteins, Signal Transducing,
pubmed-meshheading:11432792-Adult,
pubmed-meshheading:11432792-Antigens, Differentiation,
pubmed-meshheading:11432792-Brain,
pubmed-meshheading:11432792-Carrier Proteins,
pubmed-meshheading:11432792-Cell Line,
pubmed-meshheading:11432792-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:11432792-GRB2 Adaptor Protein,
pubmed-meshheading:11432792-Humans,
pubmed-meshheading:11432792-Intracellular Signaling Peptides and Proteins,
pubmed-meshheading:11432792-Lipoproteins,
pubmed-meshheading:11432792-Membrane Glycoproteins,
pubmed-meshheading:11432792-Membrane Proteins,
pubmed-meshheading:11432792-Neural Cell Adhesion Molecule L1,
pubmed-meshheading:11432792-Neural Cell Adhesion Molecules,
pubmed-meshheading:11432792-Phosphoproteins,
pubmed-meshheading:11432792-Phosphorylation,
pubmed-meshheading:11432792-Phosphotyrosine,
pubmed-meshheading:11432792-Protein Binding,
pubmed-meshheading:11432792-Protein Structure, Tertiary,
pubmed-meshheading:11432792-Protein Tyrosine Phosphatase, Non-Receptor Type 11,
pubmed-meshheading:11432792-Protein Tyrosine Phosphatase, Non-Receptor Type 6,
pubmed-meshheading:11432792-Protein Tyrosine Phosphatases,
pubmed-meshheading:11432792-Proteins,
pubmed-meshheading:11432792-Receptor, trkB,
pubmed-meshheading:11432792-Receptors, Immunologic,
pubmed-meshheading:11432792-Signal Transduction,
pubmed-meshheading:11432792-Two-Hybrid System Techniques
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pubmed:year |
2001
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pubmed:articleTitle |
Analysis of tyrosine phosphorylation-dependent protein-protein interactions in TrkB-mediated intracellular signaling using modified yeast two-hybrid system.
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pubmed:affiliation |
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan. yamada@protein.osaka-u.ac.jp
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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