Quantitative interactomics in primary T cells unveils TCR signal diversification extent and dynamics

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CD4-Positive T-Lymphocytes / immunology*
  • CD4-Positive T-Lymphocytes / metabolism
  • Chromatography, Affinity / methods
  • Mass Spectrometry / methods
  • Mice
  • Mice, Transgenic
  • Primary Cell Culture
  • Protein Interaction Mapping / methods
  • Protein Interaction Maps / immunology*
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, Antigen, T-Cell / metabolism*
  • Signal Transduction / genetics
  • Signal Transduction / immunology*


  • Receptors, Antigen, T-Cell