Figure
AP-MS workflow for analysis of the TCR signalosome. We analyze by mass spectrometry signalling complexes that form around canonical proteins used by the proximal TCR signal transduction pathway, and monitor their dynamic of assembly in the first minutes following engagement of the TCR. For that, we use genetically engineered mice models, which express at endogenous level a bait protein with a OneStreptag at their C-terminus, allowing the purification of physiological complexes, directly from primary cells, in a very efficient way. We could identify a global interaction network involving more than 200 unique proteins and more than 300 high-confidence protein-protein interactions.
Selected publications
- Reginald et al. Revisiting the Timing of Action of the PAG Adaptor Using Quantitative Proteomics Analysis of Primary T Cells. J Immunol 2015 https://doi.org/10.4049/jimmunol.1501300
- Voisinne et al. Co-recruitment analysis of the CBL and CBLB signalosomes in primary T cells identifies CD5 as a key regulator of TCR-induced ubiquitylation. Mol Syst Biol 2016 https://doi.org/10.15252/msb.20166837
- Gaud et al. The costimulatory molecule CD226 signals through VAV1 to amplify TCR signals and promote IL-17 production by CD4+T cells. Sci. Signal. 2018. https://doi.org/10.1126/scisignal.aar3083
- Voisinne et al. Quantitative interactomics in primary T cells unveils TCR signal diversification extent and dynamics. Nat. Immunol, in press