Quantitative proteomics by amino acid labeling identifies novel NHR-49 regulated proteins in C. elegans.
Stable isotope labeling by amino acids combined with mass spectrometry is a widely used methodology to quantitatively examine metabolic and signaling pathways in yeast, fruit flies, plants, cell cultures and mice. However, only metabolic labeling using (15)N has been applied to examine such events in the nematode Caenorhabditis elegans. We have recently shown that C. elegans can be completely labeled with heavy-labeled lysine by feeding worms on prelabeled lysine auxotroph Escherichia coli for just one generation. We applied this methodology to examine the organismal response to functional loss or RNAi mediated knock down of the transcription factor NHR-49, and found numerous proteins involved in lipid metabolism to be downregulated, which is consistent with its previously proposed function as a transcriptional regulator of fatty acid metabolism. The combined use of quantitative proteomics and selective gene knockdown by RNAi provides a powerful tool with broad implications for C. elegans biology.
Department of Biochemistry and Molecular Biology; University of Southern Denmark; Odense, Denmark.
Julius
J
Fredens
Nils J
NJ
Færgeman
Metabolic labeling; RNAi; SILAC; lipid metabolism; nuclear hormone receptors; proteomics; stable isotope labeling