Stable isotope labeling by amino acids combined with mass spectrometry is a widely used methodology for measuring relative changes in protein and phosphorylation levels at a global level. We have applied this method to the model organism Caenorhabditis elegans in combination with RNAi-mediated gene knockdown by feeding the nematode on pre-labeled lysine auxotroph Escherichia coli. In this chapter, we describe in details the generation of the E. coli strain, incorporation of heavy isotope-labeled lysine in C. elegans, and the procedure for a comprehensive global phosphoproteomic experiment. © 2014 Springer Science+Business Media New York.
CITATION STYLE
Fredens, J., Engholm-Keller, K., Møller-Jensen, J., Larsen, M. R., & Færgeman, N. J. (2014). Identification of novel protein functions and signaling mechanisms by genetics and quantitative phosphoproteomics in Caenorhabditis elegans. Methods in Molecular Biology, 1188, 107–124. https://doi.org/10.1007/978-1-4939-1142-4_9
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