Time-resolved phosphoproteomic analysis of the TCR signaling pathway in primary T cells

T cell receptor (TCR) signaling is essential for the function of T cells. Here we monitor and quantify the dynamics of phosphorylation sites in primary CD4+ T cells during the first 10 minutes after antibody-based TCR stimulation. To capture the earliest molecular events induced by TCR engagement we prepared unstimulated and stimulated T cells (15, 30, 120, 300 or 600 seconds after stimulation). Cells were then lysed in urea, proteins were digested into tryptic peptides, and phosphorylated peptides were enriched using titanium (TiO2) beads. In most experiments a further enrichment step was performed using phospho-Tyrosine (pTyr) immuno-precipitation. We performed 4 independent experiments, each containing 6 time-points of stimulation. This dataset contains the raw LC-MS/MS files of the pre-enriched peptide samples (SP, 24 samples) and the matching TiO2-enriched peptides (TiO2, 24 samples), as well as the eluates from further pTyr immuno-precipitation from 3 experiments (Ptyr, replicates 1, 3, 4: 18 samples). All samples were analyzed in triplicate or quadruplicate nanoLC-MS runs.