title: Global analysis of cellular protein translation by pulsed SILAC.
abstract: Current methods for system-wide gene expression analysis detect changes in mRNA abundance, but neglect regulation at the level of translation. Pulse labeling with stable isotopes has been used to measure protein turnover rates, but this does not directly provide information about translation rates. Here, we developed pulsed stable isotope labeling by amino acids in cell culture (pSILAC) with two heavy isotope labels to directly quantify protein translation on a proteome-wide scale. We applied the method to cellular iron homeostasis as a model system and demonstrate that it can confidently identify proteins that are translationally regulated by iron availability.
authors.affiliation: Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Schwanhäusser
authors.forename: Manfred
authors.initials: M
authors.lastname: Gossen
authors.forename: Gunnar
authors.initials: G
authors.lastname: Dittmar
authors.forename: Matthias
authors.initials: M
authors.lastname: Selbach
mesh_terms: D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism; D005786:Gene Expression Regulation; D015966:Gene Expression Regulation, Fungal; D006367:HeLa Cells; D006801:Humans; D007501:Iron / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods*; D008156:Luciferases / Q000032:analysis / Q000378:metabolism; D014176:Protein Biosynthesis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D015003:Yeasts
chemical_list: D000596:Amino Acids; D005656:Fungal Proteins; D020543:Proteome; D007501:Iron; D008156:Luciferases