Title: | NeuCode Labeling in Nematodes: Proteomic and Phosphoproteomic Impact of Ascaroside Treatment in Caenorhabditis elegans |
Author(s): | Rhoads TW; Prasad A; Kwiecien NW; Merrill AE; Zawack K; Westphall MS; Schroeder FC; Kimble J; Coon JJ; |
Address: | "paragraph signGenome Center. ||Biochemistry, and **Howard Hughes Medical Institute, University of Wisconsin-Madison, Madison, Wisconsin, 53706; From the Departments of double daggerChemistry, paragraph signGenome Center. double daggerdouble daggerBoyce Thompson Institute and Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York, 14853. From the Departments of double daggerChemistry, section signBiomolecular Chemistry, paragraph signGenome Center, jcoon@chem.wisc.edu" |
ISSN/ISBN: | 1535-9484 (Electronic) 1535-9476 (Print) 1535-9476 (Linking) |
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" |
Keywords: | Amino Acid Sequence Animals Caenorhabditis elegans/chemistry/*drug effects/metabolism Caenorhabditis elegans Proteins/*chemistry/isolation & purification/metabolism Escherichia coli/chemistry Food Chain Glycolipids/*pharmacology Isotope Labeling/methods L; |
Notes: | "MedlineRhoads, Timothy W Prasad, Aman Kwiecien, Nicholas W Merrill, Anna E Zawack, Kelson Westphall, Michael S Schroeder, Frank C Kimble, Judith Coon, Joshua J eng T32 GM008692/GM/NIGMS NIH HHS/ T15 LM007359/LM/NLM NIH HHS/ 5T32GM00869217/GM/NIGMS NIH HHS/ R01 GM080148/GM/NIGMS NIH HHS/ Howard Hughes Medical Institute/ R01 GM088290/GM/NIGMS NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't 2015/09/24 Mol Cell Proteomics. 2015 Nov; 14(11):2922-35. doi: 10.1074/mcp.M115.049684. Epub 2015 Sep 21" |