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      Tyrosine 1213 of Flt-1 is a major binding site of Nck and SHP-2.

      Biochemical and Biophysical Research Communications
      Adaptor Proteins, Signal Transducing, Animals, Base Sequence, Binding Sites, genetics, DNA Primers, Intracellular Signaling Peptides and Proteins, Isoenzymes, metabolism, Mutagenesis, Site-Directed, Oncogene Proteins, Phosphatidylinositol 3-Kinases, Phospholipase C gamma, Protein Tyrosine Phosphatase, Non-Receptor Type 11, Protein Tyrosine Phosphatase, Non-Receptor Type 6, Protein Tyrosine Phosphatases, Proto-Oncogene Proteins, chemistry, Receptor Protein-Tyrosine Kinases, SH2 Domain-Containing Protein Tyrosine Phosphatases, Saccharomyces cerevisiae, Signal Transduction, Transformation, Genetic, Type C Phospholipases, Tyrosine, Vascular Endothelial Growth Factor Receptor-1, src Homology Domains

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          Abstract

          Vascular endothelial growth factor (VEGF) binds to its receptor tyrosine kinase Flt-1 and KDR/Flk-1 and stimulates their autophosphorylation. However, little is known about their downstream signal transduction properties. We examined the interactions of certain proteins with a SH2-domain with Flt-1 and KDR using the yeast two-hybrid system and found that Nck, SHP-2, PLC gamma, and PI3K p85 bind to Flt-1. Extensive site-directed mutagenesis of Flt-1 revealed their major binding sites. Nck, SHP-2, and PI3K bind to Y1213 of Flt-1. Nck also binds to Y1333 of Flt-1. These results suggest that Nck, SHP-2, PLC gamma, and PI3K play important roles in Flt-1 signal transduction and that Y1213 of Flt-1 is a major binding site of PI3K, Nck, and SHP-2.

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