High-confidence glycosome proteome for procyclic form Trypanosoma brucei by epitope-tag organelle enrichment and SILAC proteomics

Maria Lucia S. Güther, Michael D. Urbaniak, Amy Tavendale, Alan Prescott, Michael A. J. Ferguson (Lead / Corresponding author)

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    Abstract

    The glycosome of the pathogenic African trypanosome Trypanosoma brucei is a specialised peroxisome that contains most of the enzymes of glycolysis and several other metabolic and catabolic pathways. The contents and transporters of this membrane-bounded organelle are of considerable interest as potential drug targets. Here, we use epitope tagging, magnetic bead enrichment and SILAC quantitative proteomics to determine a high-confidence glycosome proteome for the procyclic life cycle stage of the parasite using isotope ratios to discriminate glycosomal from mitochondrial and other contaminating proteins. The data confirm the presence of several previously demonstrated and suggested pathways in the organelle and identify previously unanticipated activities, such as protein phosphatases. The implications of the findings are discussed.

    Original languageEnglish
    Pages (from-to)2796-2806
    Number of pages11
    JournalJournal of Proteome Research
    Volume13
    Issue number6
    Early online date5 May 2014
    DOIs
    Publication statusPublished - Jun 2014

    Fingerprint

    Microbodies
    Trypanosoma brucei brucei
    Membrane Transport Proteins
    Phosphoprotein Phosphatases
    Proteome
    Isotopes
    Organelles
    Proteomics
    Life cycle
    Epitopes
    Peroxisomes
    Trypanosomiasis
    Glycolysis
    Enzymes
    Metabolic Networks and Pathways
    Life Cycle Stages
    Pharmaceutical Preparations
    Parasites
    Proteins

    Cite this

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    title = "High-confidence glycosome proteome for procyclic form Trypanosoma brucei by epitope-tag organelle enrichment and SILAC proteomics",
    abstract = "The glycosome of the pathogenic African trypanosome Trypanosoma brucei is a specialised peroxisome that contains most of the enzymes of glycolysis and several other metabolic and catabolic pathways. The contents and transporters of this membrane-bounded organelle are of considerable interest as potential drug targets. Here, we use epitope tagging, magnetic bead enrichment and SILAC quantitative proteomics to determine a high-confidence glycosome proteome for the procyclic life cycle stage of the parasite using isotope ratios to discriminate glycosomal from mitochondrial and other contaminating proteins. The data confirm the presence of several previously demonstrated and suggested pathways in the organelle and identify previously unanticipated activities, such as protein phosphatases. The implications of the findings are discussed.",
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    AU - Güther, Maria Lucia S.

    AU - Urbaniak, Michael D.

    AU - Tavendale, Amy

    AU - Prescott, Alan

    AU - Ferguson, Michael A. J.

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