Studies on fucosylation in Trypanosoma brucei

  • Giulia Bandini

    Student thesis: Doctoral ThesisDoctor of Philosophy

    Abstract

    The biosynthesis of GDP-Fucose, the activated donor for fucose, has been recently shown to be essential in the parasite Trypanosoma brucei. Fucose is a common sugar modification on eukaryotic glycan structures, but it has not been well described in trypanosomatids. To elucidate the role of fucose in T. brucei we searched for putative fucosyltransferases in this parasite. A single putative T. brucei fucosyltransferase (TbFT) was identified and recombinantly expressed in Escherichia coli. The protein was active and structural characterization of its reaction product identified it as a GDP-Fuc: ß-D-galactose a-1,2-fucosyltransferase with preference for Galß1,3GlcNAc containing structures as glycan acceptors. A procyclic form conditional null mutant for TbFT was generated and this glycosyltransferase shown to be essential for parasite growth in vitro, with the mutant cells displaying a slightly abnormal morphology and an apparent reduction in the surface high molecular weight glycoconjugate complex. Here we also describe the various experimental approaches that were used to try to identify the fucosylated glycocojugates in T. brucei. Lastly, to better understand the biosynthesis of GDP-Mannose, the starting metabolite for the biosynthesis of GDP-Fuc, we biochemically characterized T. brucei phosphomannomutase (TbPMM). Here we show this enzyme could interconvert not only mannose-phosphates, but also glucose-phosphates.
    Date of Award2011
    LanguageEnglish
    Awarding Institution
    • University of Dundee
    Add any sponsors of the thesis researchBiotechnology and Biological Sciences Research Council
    SupervisorMichael Ferguson (Supervisor)

    Keywords

    • Trypanosoma brucei
    • Fucose
    • Glycobiology

    Cite this

    Studies on fucosylation in Trypanosoma brucei
    Bandini, G. (Author). 2011

    Student thesis: Doctoral ThesisDoctor of Philosophy