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Versatile proteome labelling in fruit flies with SILAF

Florian A. Schober, Ilian Atanassov, David Moore, Anna Wedell, Christoph Freyer, Anna Wredenberg
doi: https://doi.org/10.1101/590315
Florian A. Schober
1Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm 17176, Sweden
2Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, 17165 Stockholm, Sweden
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Ilian Atanassov
3Proteomics Core Facility, Max Planck Institute for Biology of Ageing, 50931 Cologne, Germany
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  • For correspondence: ilian.atanassov@age.mpg.de christoph.freyer@ki.se anna.wredenberg@ki.se
David Moore
2Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, 17165 Stockholm, Sweden
4Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17165 Stockholm, Sweden
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Anna Wedell
1Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm 17176, Sweden
2Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, 17165 Stockholm, Sweden
5Centre for Inherited Metabolic Diseases, Karolinska University Hospital, 17176 Stockholm, Sweden
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Christoph Freyer
2Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, 17165 Stockholm, Sweden
4Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17165 Stockholm, Sweden
5Centre for Inherited Metabolic Diseases, Karolinska University Hospital, 17176 Stockholm, Sweden
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  • For correspondence: ilian.atanassov@age.mpg.de christoph.freyer@ki.se anna.wredenberg@ki.se
Anna Wredenberg
2Max Planck Institute Biology of Ageing - Karolinska Institutet Laboratory, Division of Metabolic Diseases, Department of Laboratory Medicine, Karolinska Institutet, 17165 Stockholm, Sweden
4Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 17165 Stockholm, Sweden
5Centre for Inherited Metabolic Diseases, Karolinska University Hospital, 17176 Stockholm, Sweden
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  • For correspondence: ilian.atanassov@age.mpg.de christoph.freyer@ki.se anna.wredenberg@ki.se
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ABSTRACT

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.

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Posted March 29, 2019.
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Versatile proteome labelling in fruit flies with SILAF
Florian A. Schober, Ilian Atanassov, David Moore, Anna Wedell, Christoph Freyer, Anna Wredenberg
bioRxiv 590315; doi: https://doi.org/10.1101/590315
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Versatile proteome labelling in fruit flies with SILAF
Florian A. Schober, Ilian Atanassov, David Moore, Anna Wedell, Christoph Freyer, Anna Wredenberg
bioRxiv 590315; doi: https://doi.org/10.1101/590315

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