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 specialized 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.
authors.affiliation: Division of Biological Chemistry and Drug Discovery and ‡Centre for Advanced Scientific Technologies, College of Life Sciences, University of Dundee , Dow Street, Dundee DD1 5EH, United Kingdom.
authors.forename: Maria Lucia S
authors.initials: ML
authors.lastname: Güther
authors.forename: Michael D
authors.initials: MD
authors.lastname: Urbaniak
authors.forename: Amy
authors.initials: A
authors.lastname: Tavendale
authors.forename: Alan
authors.initials: A
authors.lastname: Prescott
authors.forename: Michael A J
authors.initials: MA
authors.lastname: Ferguson
mesh_terms: D004721:Endoplasmic Reticulum / Q000378:metabolism; D000939:Epitopes; D007553:Isotope Labeling; D008018:Life Cycle Stages; D008830:Microbodies / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D015800:Protozoan Proteins / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000378:metabolism*
chemical_list: D000939:Epitopes; D024101:Mitochondrial Proteins; D020543:Proteome; D015800:Protozoan Proteins