The SILAC fly allows for accurate protein quantification in vivo.
Stable isotope labeling by amino acids in cell culture (SILAC) is widely used to quantify protein abundance in tissue culture cells. Until now, the only multicellular organism completely labeled at the amino acid level was the laboratory mouse. The fruit fly Drosophila melanogaster is one of the most widely used small animal models in biology. Here, we show that feeding flies with SILAC-labeled yeast leads to almost complete labeling in the first filial generation. We used these "SILAC flies" to investigate sexual dimorphism of protein abundance in D. melanogaster. Quantitative proteome comparison of adult male and female flies revealed distinct biological processes specific for each sex. Using a tudor mutant that is defective for germ cell generation allowed us to differentiate between sex-specific protein expression in the germ line and somatic tissue. We identified many proteins with known sex-specific expression bias. In addition, several new proteins with a potential role in sexual dimorphism were identified. Collectively, our data show that the SILAC fly can be used to accurately quantify protein abundance in vivo. The approach is simple, fast, and cost-effective, making SILAC flies an attractive model system for the emerging field of in vivo quantitative proteomics.
Sury
Matthias D
MD
Max Delbrück Center for Molecular Medicine, Robert-Rössle-Strasse 10, D-13092 Berlin, Germany.
Chen
Jia-Xuan
JX
Selbach
Matthias
M
D000818:Animals; D002853:Chromatography, Liquid; D004331:Drosophila melanogaster / Q000737:chemistry*; D005260:Female; D008239:Lysine / Q000737:chemistry; D008297:Male; D023421:Models, Animal; D010449:Peptide Mapping; D011506:Proteins / Q000032:analysis*; D053719:Tandem Mass Spectrometry
D011506:Proteins; D008239:Lysine