Lou Jieqiong, Pandžić Elvis, Böcking Till, Deng Qiji, Rossy Jérémie, Gaus Katharina
EMBL Australia Node in Single Molecule Science, School of Biomedical Sciences, University of New South Wales, Sydney, NSW 2052, Australia.
ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, NSW 2052, Australia.
Int J Mol Sci. 2025 Apr 25;26(9):4076. doi: 10.3390/ijms26094076.
Linker for activation of T cells (LAT) is an essential adaptor protein in early T cell receptor (TCR) signaling that propagates multiple signaling pathways. However, how LAT spatial organization facilitates signal initiation and propagation after TCR triggering is not clear. To differentiate de novo assembly in the plasma membrane from pre-existing LAT vesicles and clusters, we developed imaging protocols and analyses to capture the organization and dynamics of single LAT molecules immediately after TCR engagement. We could observe individual LAT molecules in the plasma membrane that assembled into immobile signaling entities requiring LAT phosphorylation. This step-wise assembly process was temporally highly coordinated via the zeta-chain-associated protein kinase 70 (Zap70)-LAT-growth factor receptor-bound protein 2 (Grb2) pathway. While multiple spatial organization co-existed even within the plasma membrane, our data suggest that de novo plasma membrane assemblies facilitated signal propagation.
T细胞活化连接蛋白（LAT）是早期T细胞受体（TCR）信号传导中一种重要的衔接蛋白，可传播多种信号通路。然而，TCR触发后LAT的空间组织如何促进信号起始和传播尚不清楚。为了区分质膜中的从头组装与预先存在的LAT囊泡和簇，我们开发了成像方案和分析方法，以捕获TCR结合后立即出现的单个LAT分子的组织和动态变化。我们能够观察到质膜中的单个LAT分子组装成需要LAT磷酸化的固定信号实体。这一逐步组装过程通过ζ链相关蛋白激酶70（Zap70）-LAT-生长因子受体结合蛋白2（Grb2）途径在时间上高度协调。虽然即使在质膜内也存在多种空间组织，但我们的数据表明，质膜的从头组装促进了信号传播。
