Interactions of Cbl with Grb2 and phosphatidylinositol 3'-kinase in activated Jurkat cells
UMass Chan Affiliations
Program in Molecular MedicineDepartment of Biochemistry and Molecular Biology
Document Type
Journal ArticlePublication Date
1995-07-01Keywords
1-Phosphatidylinositol 3-Kinase*Adaptor Proteins, Signal Transducing
Antigens, CD3
Base Sequence
Blotting, Western
GRB2 Adaptor Protein
Lymphocyte Activation
Membrane Proteins
Molecular Sequence Data
Phosphotransferases (Alcohol Group Acceptor)
Precipitin Tests
Protein Binding
Proteins
Proto-Oncogene Proteins
Proto-Oncogene Proteins c-cbl
Receptors, Antigen, T-Cell
Recombinant Fusion Proteins
*Signal Transduction
Son of Sevenless Proteins
T-Lymphocytes
*Ubiquitin-Protein Ligases
Life Sciences
Medicine and Health Sciences
Metadata
Show full item recordAbstract
T-cell receptor (TCR) cross-linking increases tyrosine phosphorylation of multiple proteins, only a few of which have been identified. One of the most rapidly tyrosine-phosphorylated polypeptides is the 120-kDa product of the proto-oncogene c-cbl, a cytosolic and cytoskeletal protein containing multiple proline-rich motifs that are potential binding sites for proteins containing Src homology 3 (SH3) domains. We report here that in cultured Jurkat T cells, Cbl is coprecipitated with antibody against the adapter protein Grb2. Upon activation of Jurkat T cells via the TCR-CD3 complex, we find that high-affinity binding of Cbl requires the N-terminal SH3 domain of GST-Grb2 fusion protein but after cross-linking of the TCR-CD3 and CD4 receptors, Cbl binds equally to its SH2 domain. Grb2 antisera also precipitated p85 from serum-starved cells, while TCR activation increased p85 and tyrosine-phosphorylated Cbl but not Cbl protein in Grb2 immunocomplexes. Phosphatidylinositol (PI) 3-kinase activity was immunoprecipitated from serum-starved cells with Cbl and to a lesser extent with Grb2 antisera, and TCR cross-linking increased this activity severalfold. The PI 3-kinase activity associated with Cbl amounted to 5 to 10% of the total cellular activity that could be precipitated by p85 antisera. The Ras exchange factor Son-of-sevenless 1 (Sos-1) was not found in anti-Cbl immunoprecipitates from activated cells, and Cbl was not detectable in anti-Sos-1 precipitates, supporting the likelihood that Sos-Grb2 and Cbl-Grb2 are present as distinct complexes. Taken together, these data suggest that Cbl function in Jurkat T cells involves its constitutive association with Grb2 and its recruitment of PI 3-kinase in response to TCR activation.Source
Mol Cell Biol. 1995 Jul;15(7):3571-8.Permanent Link to this Item
http://hdl.handle.net/20.500.14038/38592PubMed ID
7791764Related Resources
Link to Article in PubMedCollections
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