Global analysis of cellular protein translation by pulsed SILAC

作者：Björn Schwanhäußer, Manfred Gossen, Gunnar Dittmar, Matthias Selbach · 发表于：PROTEOMICS · 年份：2008 · DOI：10.1002/pmic.200800275 · 被引用次数：375 · 研究领域：RNA and protein synthesis mechanisms、Advanced Proteomics Techniques and Applications、Mitochondrial Function and Pathology

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.