1. Global analysis of cellular protein translation by pulsed SILAC
- Author
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Björn Schwanhäusser, Matthias Selbach, Gunnar Dittmar, and Manfred Gossen
- Subjects
Proteomics ,Proteome ,Iron ,Biology ,Biochemistry ,Fungal Proteins ,Gene Expression Regulation, Fungal ,Yeasts ,Stable isotope labeling by amino acids in cell culture ,Translational regulation ,Protein biosynthesis ,Animals ,Humans ,Amino Acids ,Luciferases ,Molecular Biology ,Cells, Cultured ,Fungal protein ,Stable isotope ratio ,Protein turnover ,Translation (biology) ,Genetic translation ,Cell biology ,Gene Expression Regulation ,Isotope Labeling ,Protein Biosynthesis ,HeLa Cells - 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.
- Published
- 2009
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