Identification of two tyrosine phosphoproteins, pp70 and pp68, which interact with phospholipase Cgamma, Grb2, and Vav after B cell antigen receptor activation.
Tyrosine phosphorylation of cellular proteins mediates the assembly and localization of effector proteins through interactions facilitated by modular Src homology 2 (SH2) and phosphotyrosine binding domains. We describe here two tyrosine-phosphorylated proteins with Mr values of 70,000 and 68,000 that interact with Grb2, phospholipase C (PLCgamma1 and PLCgamma2), and Vav after B cell receptor cross-linking. The interaction of pp70 and pp68 with PLC and Vav is mediated by the carboxyl-terminal SH2 domain of PLC and the SH2 domain of Vav. In contrast, the interaction of pp70 and pp68 with Grb2 requires cooperative binding of the SH2 and SH3 domains of Grb2. Western blot analysis demonstrated that neither pp70 nor pp68 represented the recently described linker protein SLP-76, which binds Grb2, PLC, and Vav in T cells after T cell receptor activation. Moreover, SLP-76 protein was not detected in a number of B cell lines or in normal mouse B cells. Hence, we propose that pp70 and pp68 likely represent B cell homologs of SLP-76 which facilitate and coordinate B cell activation.
Fu
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Program in Genetics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Chan
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D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology*; D001665:Binding Sites; D002051:Burkitt Lymphoma; D018797:Cell Cycle Proteins; D002460:Cell Line; D066246:ErbB Receptors / Q000378:metabolism; D051380:GRB2 Adaptor Protein; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D051966:Phospholipase C gamma; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism*; D019000:Phosphotyrosine; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051567:Proto-Oncogene Proteins c-vav; D011947:Receptors, Antigen, B-Cell / Q000502:physiology*; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000302:isolation & purification / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; C495949:Grb2 protein, mouse; D007527:Isoenzymes; D010750:Phosphoproteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D051567:Proto-Oncogene Proteins c-vav; D011947:Receptors, Antigen, B-Cell; C496618:VAV1 protein, human; C496619:Vav1 protein, mouse; D019000:Phosphotyrosine; D066246:ErbB Receptors; D010738:Type C Phospholipases; D051966:Phospholipase C gamma