Multiple SH2-mediated interactions in v-src-transformed cells.
The Src homology 2 (SH2) domain is a noncatalytic region which is conserved among a number of signaling and transforming proteins, including cytoplasmic protein-tyrosine kinases and Ras GTPase-activating protein (GAP). Genetic and biochemical data indicate that the SH2 domain of the p60v-src (v-Src) protein-tyrosine kinase is required for full v-src ... transforming activity and may direct the association of v-Src with specific tyrosine-phosphorylated proteins. To test the ability of the v-Src SH2 domain to mediate protein-protein interactions, v-Src polypeptides were expressed as fusion proteins in Escherichia coli. The bacterial v-Src SH2 domain bound a series of tyrosine-phosphorylated proteins in a lysate of v-src-transformed Rat-2 cells, including prominent species of 130 and 62 kDa (p130 and p62). The p130 and p62 tyrosine-phosphorylated proteins that complexed v-Src SH2 in vitro also associated with v-Src in v-src-transformed Rat-2 cells; this in vivo binding was dependent on the v-Src SH2 domain. In addition to binding soluble p62 and p130, the SH2 domains of v-Src, GAP, and v-Crk directly recognized these phosphotyrosine-containing proteins which had been previously denatured and immobilized on a filter. In addition, the SH2 domains of GAP and v-Crk bound to the GAP-associated protein p190 immobilized on a nitrocellulose membrane. These results show that SH2 domains bind directly to tyrosine-phosphorylated proteins and that the Src SH2 domain can bind phosphorylated targets of the v-Src kinase domain.(ABSTRACT TRUNCATED AT 250 WORDS)
Mesh Terms:
Animals, Cell Line, Transformed, Cell Transformation, Neoplastic, Escherichia coli, GTPase-Activating Proteins, Oncogene Protein pp60(v-src), Peptide Mapping, Phosphorylation, Protein-Tyrosine Kinases, Proteins, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-crk, Rats, Recombinant Fusion Proteins, Sequence Homology, Nucleic Acid, Tyrosine, ras GTPase-Activating Proteins
Animals, Cell Line, Transformed, Cell Transformation, Neoplastic, Escherichia coli, GTPase-Activating Proteins, Oncogene Protein pp60(v-src), Peptide Mapping, Phosphorylation, Protein-Tyrosine Kinases, Proteins, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-crk, Rats, Recombinant Fusion Proteins, Sequence Homology, Nucleic Acid, Tyrosine, ras GTPase-Activating Proteins
Mol. Cell. Biol.
Date: Mar. 01, 1992
PubMed ID: 1545818
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