Identification of amino acid residues required for a specific interaction between Src-tyrosine kinase and proline-rich region of phosphatidylinositol-3' kinase.
The binding of ligand to B-cell antigen receptors (BCR) leads to the activation of receptor-associated Src-family kinases and phosphatidylinositol-3' kinase (PI-3 kinase). Although it has been demonstrated that SH3 domains of several Src-family kinases interact with PI-3 kinase by binding to a proline-rich region of PI-3 kinase in vitro, there is no direct evidence to support their interaction in vivo. Thus, we utilized the yeast two-hybrid assay to reconstitute this protein-protein interaction. This genetic screen clearly indicates that the interaction between SH3 domain of Fyn and the proline-rich region (residues: 80-104) of PI-3 kinase is highly specific. Mutational analysis revealed that amino acid residues Asp92, Tyr93, Arg96 and Thr97 of the SH3 domain of Fyn are essential for interacting with the proline-rich peptide of PI-3 kinase.
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Wyeth-Ayerst Research, Molecular Biology Section, Pearl River, NY 10965, USA.
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Kurosaki
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D000595:Amino Acid Sequence; D003470:Culture Media; D008969:Molecular Sequence Data; D009154:Mutation; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000378:metabolism*; D051076:Proto-Oncogene Proteins c-fyn; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic; D018909:src Homology Domains
D003470:Culture Media; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D051076:Proto-Oncogene Proteins c-fyn; D011993:Recombinant Fusion Proteins