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- Title
Metabolic labeling of C. elegans and D. melanogaster for quantitative proteomics.
- Authors
Krijgsveld, Jeroen; Ketting, René F; Mahmoudi, Tokameh; Johansen, Janik; Artal-Sanz, Marta; Verrijzer, C Peter; Plasterk, Ronald H A; Heck, Albert J R
- Abstract
A crucial issue in comparative proteomics is the accurate quantification of differences in protein expression levels. To achieve this, several methods have been developed in which proteins are labeled with stable isotopes eitherin vivo via metabolic labeling orin vitro by protein derivatization. Although metabolic labeling is the only way to obtain labeling of all proteins, it has thus far only been applied to single-celled organisms[SUP1,2] and cells in culture[SUP(2,3]. Here we describe quantitative[SUP15] N metabolic labeling of the multicellular organismsCaenorhabditis elegans, a nematode, and Drosophila melanogaster, the common fruit fly, achieved by feeding them on[SUP15] N-labeledEscherichia coli and yeast, respectively. The relative abundance of individual proteins obtained from different samples can then be determined by mass spectrometry (MS). The applicability of the method is exemplified by the comparison of protein expression levels in two C. elegans strains, one with and one without a germ line. The methodology described provides tools for accurate quantitative proteomic studies in these model organisms.
- Subjects
PROTEOMICS; DERIVATIZATION; CAENORHABDITIS elegans; MASS spectrometry
- Publication
Nature Biotechnology, 2003, Vol 21, Issue 8, p927
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt848