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NeuCode Labeling in Nematodes: Proteomic and Phosphoproteomic Impact of Ascaroside Treatment in Caenorhabditis elegans
Journal article   Open access   Peer reviewed

NeuCode Labeling in Nematodes: Proteomic and Phosphoproteomic Impact of Ascaroside Treatment in Caenorhabditis elegans

Timothy W Rhoads, Aman Prasad, Nicholas W Kwiecien, Anna E Merrill, Kelson Zawack, Michael S Westphall, Frank C Schroeder, Judith Kimble and Joshua J Coon
Molecular & cellular proteomics, Vol.14(11), pp.2922-2935
11/2015
DOI: 10.1074/mcp.M115.049684
PMCID: PMC4638036
PMID: 26392051
url
https://doi.org/10.1074/mcp.M115.049684View
Published (Version of record) Open Access

Abstract

The nematode Caenorhabditis elegans is an important model organism for biomedical research. We previously described NeuCode stable isotope labeling by amino acids in cell culture (SILAC), a method for accurate proteome quantification with potential for multiplexing beyond the limits of traditional stable isotope labeling by amino acids in cell culture. Here we apply NeuCode SILAC to profile the proteomic and phosphoproteomic response of C. elegans to two potent members of the ascaroside family of nematode pheromones. By consuming labeled E. coli as part of their diet, C. elegans nematodes quickly and easily incorporate the NeuCode heavy lysine isotopologues by the young adult stage. Using this approach, we report, at high confidence, one of the largest proteomic and phosphoproteomic data sets to date in C. elegans: 6596 proteins at a false discovery rate ≤ 1% and 6620 phosphorylation isoforms with localization probability ≥75%. Our data reveal a post-translational signature of pheromone sensing that includes many conserved proteins implicated in longevity and response to stress.
Caenorhabditis elegans - chemistry Phosphorylation Caenorhabditis elegans Proteins - chemistry Caenorhabditis elegans Proteins - metabolism Molecular Sequence Data Pheromones - isolation & purification Phosphoproteins - metabolism Proteome - chemistry Phosphoproteins - chemistry Lysine - metabolism Proteomics - methods Proteome - isolation & purification Amino Acid Sequence Caenorhabditis elegans - metabolism Glycolipids - pharmacology Caenorhabditis elegans Proteins - isolation & purification Pheromones - metabolism Escherichia coli - chemistry Protein Interaction Mapping Food Chain Isotope Labeling - methods Animals Caenorhabditis elegans - drug effects Pheromones - chemistry Phosphoproteins - isolation & purification Protein Processing, Post-Translational Lysine - chemistry Proteome - metabolism

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