QuaNCAT: quantitating proteome dynamics in primary cells.
Howden AJM., Geoghegan V., Katsch K., Efstathiou G., Bhushan B., Boutureira O., Thomas B., Trudgian DC., Kessler BM., Dieterich DC., Davis BG., Acuto O.
Here we demonstrate quantitation of stimuli-induced proteome dynamics in primary cells by combining the power of bio-orthogonal noncanonical amino acid tagging (BONCAT) and stable-isotope labeling of amino acids in cell culture (SILAC). In conjunction with nanoscale liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS), quantitative noncanonical amino acid tagging (QuaNCAT) allowed us to monitor the early expression changes of >600 proteins in primary resting T cells subjected to activation stimuli.