title: Proteomics meets genetics: SILAC labeling of Drosophila melanogaster larvae and cells for in vivo functional studies.
abstract: Stable isotope labeling by amino acids in cell culture (SILAC) is an established and potent method for quantitative proteomics. When combined with high-resolution mass spectrometry (MS) and efficient algorithms for the analysis of quantitative MS data, SILAC has proven to be the strategy of choice for the in-depth characterization of functional states at the protein level. The fruit fly Drosophila melanogaster is one of the most widely used model systems for studies of genetics and developmental biology. Despite this, a global proteomic approach in Drosophila is rarely considered. Here, we describe an adaptation of SILAC for functional investigation of fruit flies by proteomics: We illustrate how to perform efficient SILAC labeling of cells in culture and whole fly larvae. The combination of SILAC, a highly accurate global protein quantification method, and of the fruit fly, the prime genetics and developmental model, represents a unique opportunity for quantitative proteomic studies in vivo.
authors: Department of Molecular Oncology, European Institute of Oncology, Via Adamello 16, 20139, Milan, Italy.
authors: Alessandro
authors: A
authors: Cuomo
authors: Roberta
authors: R
authors: Sanfilippo
authors: Thomas
authors: T
authors: Vaccari
authors: Tiziana
authors: T
authors: Bonaldi
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D053000:Analytic Sample Preparation Methods; D000818:Animals; D002458:Cell Fractionation; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D003470:Culture Media; D004331:Drosophila melanogaster / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004625:Embryo, Nonmammalian / Q000166:cytology; D019476:Insect Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D007553:Isotope Labeling / Q000379:methods*; D007814:Larva / Q000378:metabolism; D013058:Mass Spectrometry; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D020858:Tissue and Organ Harvesting
chemical_list: D000596:Amino Acids; D003470:Culture Media; D019476:Insect Proteins