Analysis of the carbohydrate components of glycosylphosphatidylinositol structures using fluorescent labeling

Nicole Zitzmann, Michael A. J. Ferguson

    Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

    9 Citations (Scopus)

    Abstract

    Glycosylphosphatidylinositols (GPIs) are a family of structures that contain the structural motif: Mana1-4GlcNH2a1-6myo-Inositol-1-PO4-lipid. This common substructure suggests that this family of molecules is biosynthetically related, and it differentiates them from other glycosylated phosphoinositides, such as the glycosylated phosphatidylinositols of mycobacteria and the glycosylated inositol phosphoceramides of yeasts and plants. The GPI family can be conveniently divided into two groups, based on structural homology and function. The first group (1–34) is the membrane protein anchors (Fig. 1) which are found covalently linked to the C-termini of a wide variety of externally disposed plasma-membrane proteins throughout the eukaryotes. These GPI anchors afford a stable attachment of proteins to the membrane, and can be viewed as an alternative mechanism of membrane attachment to a singlepass hydrophobic transmembrane peptide domain. For recent reviews of GPI anchor structure, biosynthesis, and function, see refs. 35–40. The second group of GPI structures have only been found in protozoan organisms. These molecules exist as free glycophospholipids, such as the glycoinositol phospholipids (GIPLs) of the Leishmania, Trypanosoma cruzi, Leptomonas, Herpetomonas, Phytomonas, and Toxoplasma (35,41–45), or are attached to phosphorylated repeating units, as in the lipophosphoglycans (LPGs) of the Leishmania (35,46). This chapter describes protocols specifically designed to analyze the carbohydrate components of protein-linked GPI anchors, although they are also applicable to the GIPLs and, to some extent, the LPGs.
    Original languageEnglish
    Title of host publicationProtein lipidation protocols
    EditorsMichael H. Gelb
    Place of PublicationTotowa
    PublisherHumana Press
    Pages73-89
    Number of pages17
    ISBN (Electronic)9781592592647
    ISBN (Print)9780896035348
    DOIs
    Publication statusPublished - 1999

    Publication series

    NameMethods in Molecular Biology
    PublisherHumana Press
    Volume116
    ISSN (Print)1064-3745

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