Rapid temporal dynamics of transcription, protein synthesis, and secretion during macrophage activation.

ABSTRACT: Macrophages provide the first line of host defense with their capacity to react to an array of cytokines and bacterial components requiring tight regulation of protein expression and secretion to invoke a properly tuned innate immune response. To capture the dynamics of this system, we introduce a novel method combining pulsed stable isotope labeling with amino acids in cell culture (SILAC) with pulse labeling using the methionine analog azidohomoalanine that allows the enrichment of newly synthesized proteins via click-chemistry followed by their identification and quantification by mass spectrometry. We show that this permits the analysis of proteome changes on a rapid time scale, as evidenced by the detection of 4852 newly synthesized proteins after only a 20-min SILAC pulse. We have ap

SUBMITTER: Eichelbaum K 

PROVIDER: S-EPMC3945909 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature