Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, INSERM, CNRS, Marseille, France.
Institut de Pharmacologie et de Biologie Structurale, Département Biologie Structurale Biophysique, Protéomique Génopole Toulouse Midi Pyrénées CNRS UMR 5089, Toulouse, France.
Nat Immunol. 2019 Nov;20(11):1530-1541. doi: 10.1038/s41590-019-0489-8. Epub 2019 Oct 7.
The activation of T cells by the T cell antigen receptor (TCR) results in the formation of signaling protein complexes (signalosomes), the composition of which has not been analyzed at a systems level. Here, we isolated primary CD4 T cells from 15 gene-targeted mice, each expressing one tagged form of a canonical protein of the TCR-signaling pathway. Using affinity purification coupled with mass spectrometry, we analyzed the composition and dynamics of the signalosomes assembling around each of the tagged proteins over 600 s of TCR engagement. We showed that the TCR signal-transduction network comprises at least 277 unique proteins involved in 366 high-confidence interactions, and that TCR signals diversify extensively at the level of the plasma membrane. Integrating the cellular abundance of the interacting proteins and their interaction stoichiometry provided a quantitative and contextual view of each documented interaction, permitting anticipation of whether ablation of a single interacting protein can impinge on the whole TCR signal-transduction network.
T 细胞抗原受体 (TCR) 激活 T 细胞会导致信号蛋白复合物 (signalosomes) 的形成，但其系统水平的组成尚未得到分析。在这里，我们从 15 只基因靶向小鼠中分离出原代 CD4 T 细胞，每只小鼠表达一种 TCR 信号通路的典型蛋白的标记形式。我们使用亲和纯化结合质谱分析，分析了在 TCR 结合的 600 秒内，围绕每个标记蛋白组装的信号体的组成和动态。我们表明，TCR 信号转导网络至少包含 277 种独特的蛋白质，涉及 366 种高可信度的相互作用，并且 TCR 信号在质膜水平上广泛多样化。整合相互作用蛋白的细胞丰度及其相互作用化学计量学，为每个记录的相互作用提供了定量和上下文的视角，从而可以预测是否消除单个相互作用蛋白会影响整个 TCR 信号转导网络。
