biostudies-literatureRothenberg DASwiss National Science FoundationNIEHS NIH HHSNCI NIH HHSNIHNational Research Foundation of SingaporeMIT Center for Precision Cancer Medicine367-381https://www.ebi.ac.uk/biostudies/studies/S-EPMC6249402biostudies-literatureUnknown9To quantify dynamic protein synthesis rates, we developed MITNCAT, a method combining multiplexed isobaric mass tagging with pulsed SILAC (pSILAC) and bio-orthogonal non-canonical amino acid tagging (BONCAT) to label newly synthesized proteins with azidohomoalanine (Aha), thus enabling high temporal resolution across multiple conditions in a single analysis. MITNCAT quantification of protein synthesis rates following induction of the unfolded protein response revealed global down-regulation of protein synthesis, with stronger down-regulation of glycolytic and protein synthesis machinery proteins, but up-regulation of several key chaperones. Waves of temporally distinct protein synthesis were observed in response to epidermal growth factor, with altered synthesis detectable in the first 15 iScienceA Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth.PMC6249402CA014051ES024615CA096504P01 CA026731P30 ES002109CA210180ES026856ES002109U54 CA210180P30 CA014051P2SKP3_174681CA026731R01 CA096504R01 ES026856174681P42 ES027707R01 ES024615White FMRothenberg DAHuber SMBegley TJDedon PCTaliaferro JMfalseA Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth.To quantify dynamic protein synthesis rates, we developed MITNCAT, a method combining multiplexed isobaric mass tagging with pulsed SILAC (pSILAC) and bio-orthogonal non-canonical amino acid tagging (BONCAT) to label newly synthesized proteins with azidohomoalanine (Aha), thus enabling high temporal resolution across multiple conditions in a single analysis. MITNCAT quantification of protein synthesis rates following induction of the unfolded protein response revealed global down-regulation of protein synthesis, with stronger down-regulation of glycolytic and protein synthesis machinery proteins, but up-regulation of several key chaperones. Waves of temporally distinct protein synthesis were observed in response to epidermal growth factor, with altered synthesis detectable in the first 15 2018-01-01T00:00:00Z2018 Nov2025-04-05T12:59:46.586Z2019-03-27T00:10:00ZS-EPMC62494023046606310.1016/j.isci.2018.11.004