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Proteome-Wide Quantitative Phosphoproteomic Analysis of Trypanosoma brucei Insect and Mammalian Life Cycle Stages

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Trypanosomatids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2116))

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Abstract

Mass spectrometry based proteomics allows for the identification and quantification of protein and phosphorylation site abundance on a proteome wide scale. Here we describe the metabolic labeling of cultured Trypanosoma brucei cells in either the bloodstream or procyclic life cycle stage using stable isotope labeling of amino acids in cell culture (SILAC), and the production of samples suitable for analysis by liquid chromatography tandem mass spectrometry. The protocols require little specialist equipment, and they typically enable quantification of over 4500 proteins and 9000 phosphorylation sites.

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Authors and Affiliations

  1. Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancaster, UK

    Corinna Benz & Michael D. Urbaniak

Authors
  1. Corinna Benz
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  2. Michael D. Urbaniak
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Corresponding author

Correspondence to Michael D. Urbaniak .

Editor information

Editors and Affiliations

  1. School of Biological Sciences, University of Edinburgh, Edinburgh, UK

    Paul A. M. Michels

  2. School of Applied Sciences, University of Huddersfield, Huddersfield, UK

    Michael L. Ginger

  3. Faculty of Biology, Technion–Israel Institute of Technology, Haifa, Israel

    Dan Zilberstein

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Benz, C., Urbaniak, M.D. (2020). Proteome-Wide Quantitative Phosphoproteomic Analysis of Trypanosoma brucei Insect and Mammalian Life Cycle Stages. In: Michels, P., Ginger, M., Zilberstein, D. (eds) Trypanosomatids. Methods in Molecular Biology, vol 2116. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0294-2_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0294-2_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0293-5

  • Online ISBN: 978-1-0716-0294-2

  • eBook Packages: Springer Protocols

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