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.
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