Houtman Jon C D, Yamaguchi Hiroshi, Barda-Saad Mira, Braiman Alex, Bowden Brent, Appella Ettore, Schuck Peter, Samelson Lawrence E
Laboratory of Cellular and Molecular Biology, National Cancer Institute, US National Institutes of Health, Bethesda, Maryland 20892, USA.
Nat Struct Mol Biol. 2006 Sep;13(9):798-805. doi: 10.1038/nsmb1133. Epub 2006 Aug 13.
Receptor oligomerization is vital for activating intracellular signaling, in part by initiating events that recruit effector and adaptor proteins to sites of active signaling. Whether these distal molecules themselves oligomerize is not well appreciated. In this study, we examined the molecular interactions of the adaptor protein GRB2. In T cells, the SH2 domain of GRB2 binds phosphorylated tyrosines on the adaptor protein LAT and the GRB2 SH3 domains associate with the proline-rich regions of SOS1 and CBL. Using biochemical and biophysical techniques in conjunction with confocal microscopy, we observed that the simultaneous association of GRB2, via its SH2 and SH3 domains, with multivalent ligands led to the oligomerization of these ligands, which affected signaling. These data suggest that multipoint binding of distal adaptor proteins mediates the formation of oligomeric signaling clusters vital for intracellular signaling.
受体寡聚化对于激活细胞内信号传导至关重要,部分原因是通过启动将效应蛋白和衔接蛋白招募到活跃信号位点的事件。这些远端分子自身是否寡聚化尚未得到充分认识。在本研究中,我们检测了衔接蛋白GRB2的分子相互作用。在T细胞中,GRB2的SH2结构域结合衔接蛋白LAT上的磷酸化酪氨酸,GRB2的SH3结构域与SOS1和CBL富含脯氨酸的区域结合。结合共聚焦显微镜使用生化和生物物理技术,我们观察到GRB2通过其SH2和SH3结构域与多价配体的同时结合导致这些配体寡聚化,这影响了信号传导。这些数据表明,远端衔接蛋白的多点结合介导了对细胞内信号传导至关重要的寡聚信号簇的形成。