Versatile proteome labelling in fruit flies with SILAF

Drosophila melanogaster has been a working horse of genetics and cell biology for more than a century. However, proteomic-based methods have been limited due to technical obstacles, especially the lack of reliable labelling methods. Here, we advanced a chemically defined food source into stable-isot...

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Bibliographic Details
Published inbioRxiv
Main Authors Schober, Florian A, Atanassov, Ilian, Moore, David, Wedell, Anna, Freyer, Christoph, Wredenberg, Anna
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 29.03.2019
Subjects
Amino acids
Food sources
Labeling
Lysine
Metabolism
Phosphorylation
Post-translation
Protein turnover
Proteomes
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Summary:Drosophila melanogaster has been a working horse of genetics and cell biology for more than a century. However, proteomic-based methods have been limited due to technical obstacles, especially the lack of reliable labelling methods. Here, we advanced a chemically defined food source into stable-isotope labelling of amino acids in flies (SILAF). It allows for the rapid generation of a large number of flies with full incorporation of lysine-6. SILAF followed by fractionation and enrichment gave proteomic insights at a depth of 5,966 proteins and 7,496 phosphorylation sites, which substantiated metabolic regulation on enzymatic level. Furthermore, the label can be traced and predicts protein turnover rates during different developmental stages. The ease and versatility of the method actuates the fruit fly as an appealing model in proteomic and post-translational modification studies and it enlarges potential metabolic applications based on heavy amino acid diets.
DOI:10.1101/590315