Association of Grb-2 and PI3K p85 with phosphotyrosile peptides derived from BTLA.
B and T lymphocyte attenuator (BTLA) is a recently identified inhibitory receptor expressed by B and T cells. We previously identified two tyrosine-containing signaling motifs in the cytoplasmic domain of BTLA that interact with the SHP-1 and SHP-2 phosphatases. BTLA has a third conserved tyrosine-containing motif within the cytoplasmic domain, ... similar in sequence to a Grb-2 recruitment site. To identify specific interacting proteins that would be recruited to this motif, we carried out an unbiased screen by using synthetic peptides in active (e.g., phosphotyrosil-containing) or control (e.g., non-phosphorylated) forms as baits. Using mass spectrometry, we identified two specific interacting proteins, Grb-2 and the p85 subunit of PI3K. Further, we demonstrate that the interaction with Grb-2 is direct, whereas the recruitment of the p85 subunit by BTLA phosphotyrosile-containing peptides may be indirect via its association with Grb-2. These findings may provide biochemical basis for previously unexplained actions of BTLA.
Mesh Terms:
Amino Acid Sequence, Animals, Binding Sites, Blotting, Western, Cell Line, Tumor, GRB2 Adaptor Protein, Glutathione Transferase, Humans, Immunoprecipitation, Mice, Molecular Sequence Data, Oligopeptides, Phosphatidylinositol 3-Kinases, Phosphorylation, Phosphotyrosine, Protein Binding, Protein Subunits, Receptors, Immunologic, Recombinant Fusion Proteins, Sequence Homology, Amino Acid
Amino Acid Sequence, Animals, Binding Sites, Blotting, Western, Cell Line, Tumor, GRB2 Adaptor Protein, Glutathione Transferase, Humans, Immunoprecipitation, Mice, Molecular Sequence Data, Oligopeptides, Phosphatidylinositol 3-Kinases, Phosphorylation, Phosphotyrosine, Protein Binding, Protein Subunits, Receptors, Immunologic, Recombinant Fusion Proteins, Sequence Homology, Amino Acid
Biochem. Biophys. Res. Commun.
Date: Jul. 14, 2006
PubMed ID: 16725108
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