Analysis of the interactome of the RLTPR/CARMIL2 protein in CD4+ T cells

We analyzed the interactome of the mouse protein RLTPR (CARMIL2) using an affinity purification method based on the introduction of a One-Strep-tag (OST) sequence in the C-terminus of the bait protein by genetic engineering of primary mouse T cells. We used mouse models expressing the tagged version of either the wild-type protein or the protein bearing the Q575E mutation. Affinity purification of these OST tagged proteins was performed using Streptactin beads, from T cells either non stimulated, stimulated 2min, 5min or 10min with pervanadate. Each AP-MS purification of an OST- protein is associated with a corresponding control (purification from WT CD4+ T cells) in the same conditions of stimulation. Three replicate biological experiments were performed.