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

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

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Authors

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

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

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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