Post-translational modifications of the Dictyostelium discoideum glycoprotein PsA

glycosylphosphatidylinositol membrane anchor and composition of O-linked oligosaccharides

Paul A. Haynes, Andrew A. Gooley, Michael A. J. Ferguson, John W. Redmond, Keith L. Williams

    Research output: Contribution to journalArticle

    51 Citations (Scopus)

    Abstract

    Prespore-specific antigen (PsA) is a cell-surface glycoprotein isolated from Dictyostelium discoideum, which is post-translationally modified by addition of carbohydrate to threonine residues of the carboxy-terminal peptide domain, and a glycosylphosphatidylinositol (GPI) anchor which attaches the glycoprotein to the cell membrane. The GPI anchor was isolated by proteolytic cleavage of the protein, and the structure of the lipid and glycan portions of the anchor were determined. The lipid moiety of the anchor is an inositolphosphoceramide which contains C18:0 phytosphingosine as a long chain base, and a mixture of fatty acids with a C18:1 mono-unsaturated fatty acid as the major component. The purified GPI anchor was susceptible to digestion by a bacterial phosphatidylinositol-specific phospholipase-C enzyme. The glycan of the GPI anchor consisted of two molecular species present in the ratio 55:45, the structures of which were determined by exoglycosidase sequencing and found to be Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2 and Man alpha 1-2Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2. The glucosamine in both structures is glycosidically linked to the inositol ring of the inositolphosphoceramide. The GPI glycan structures are consistent with the conserved core structure of all characterised GPI anchors, and the structure of the D. discoideum GPI moiety has features in common with structures from yeast, protozoa and higher eukaryotes. Compositional analysis of the carbohydrate attached to threonine residues in the carboxy-terminal peptide domain is also presented. The oligosaccharides bind to wheat germ agglutinin, and contain glucosamine and fucose as the major constituents.
    Original languageEnglish
    Pages (from-to)729-737
    Number of pages9
    JournalEuropean Journal of Biochemistry
    Volume216
    Issue number3
    DOIs
    Publication statusPublished - 1993

    Fingerprint

    Glycosylphosphatidylinositols
    Post Translational Protein Processing
    Anchors
    Oligosaccharides
    Glycoproteins
    Membranes
    Antigens
    Chemical analysis
    Polysaccharides
    phytosphingosine
    Dictyostelium
    Glucosamine
    Threonine
    Monounsaturated fatty acids
    Carbohydrates
    Protozoa
    Phosphoinositide Phospholipase C
    Lipids
    Peptides
    Wheat Germ Agglutinins

    Cite this

    @article{957f1af6a67d4c84b7c6e46527051cd6,
    title = "Post-translational modifications of the Dictyostelium discoideum glycoprotein PsA: glycosylphosphatidylinositol membrane anchor and composition of O-linked oligosaccharides",
    abstract = "Prespore-specific antigen (PsA) is a cell-surface glycoprotein isolated from Dictyostelium discoideum, which is post-translationally modified by addition of carbohydrate to threonine residues of the carboxy-terminal peptide domain, and a glycosylphosphatidylinositol (GPI) anchor which attaches the glycoprotein to the cell membrane. The GPI anchor was isolated by proteolytic cleavage of the protein, and the structure of the lipid and glycan portions of the anchor were determined. The lipid moiety of the anchor is an inositolphosphoceramide which contains C18:0 phytosphingosine as a long chain base, and a mixture of fatty acids with a C18:1 mono-unsaturated fatty acid as the major component. The purified GPI anchor was susceptible to digestion by a bacterial phosphatidylinositol-specific phospholipase-C enzyme. The glycan of the GPI anchor consisted of two molecular species present in the ratio 55:45, the structures of which were determined by exoglycosidase sequencing and found to be Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2 and Man alpha 1-2Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2. The glucosamine in both structures is glycosidically linked to the inositol ring of the inositolphosphoceramide. The GPI glycan structures are consistent with the conserved core structure of all characterised GPI anchors, and the structure of the D. discoideum GPI moiety has features in common with structures from yeast, protozoa and higher eukaryotes. Compositional analysis of the carbohydrate attached to threonine residues in the carboxy-terminal peptide domain is also presented. The oligosaccharides bind to wheat germ agglutinin, and contain glucosamine and fucose as the major constituents.",
    author = "Haynes, {Paul A.} and Gooley, {Andrew A.} and Ferguson, {Michael A. J.} and Redmond, {John W.} and Williams, {Keith L.}",
    year = "1993",
    doi = "10.1111/j.1432-1033.1993.tb18192.x",
    language = "English",
    volume = "216",
    pages = "729--737",
    journal = "European Journal of Biochemistry",
    issn = "0014-2956",
    publisher = "Wiley",
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    }

    Post-translational modifications of the Dictyostelium discoideum glycoprotein PsA : glycosylphosphatidylinositol membrane anchor and composition of O-linked oligosaccharides. / Haynes, Paul A.; Gooley, Andrew A.; Ferguson, Michael A. J.; Redmond, John W.; Williams, Keith L.

    In: European Journal of Biochemistry, Vol. 216, No. 3, 1993, p. 729-737.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Post-translational modifications of the Dictyostelium discoideum glycoprotein PsA

    T2 - glycosylphosphatidylinositol membrane anchor and composition of O-linked oligosaccharides

    AU - Haynes, Paul A.

    AU - Gooley, Andrew A.

    AU - Ferguson, Michael A. J.

    AU - Redmond, John W.

    AU - Williams, Keith L.

    PY - 1993

    Y1 - 1993

    N2 - Prespore-specific antigen (PsA) is a cell-surface glycoprotein isolated from Dictyostelium discoideum, which is post-translationally modified by addition of carbohydrate to threonine residues of the carboxy-terminal peptide domain, and a glycosylphosphatidylinositol (GPI) anchor which attaches the glycoprotein to the cell membrane. The GPI anchor was isolated by proteolytic cleavage of the protein, and the structure of the lipid and glycan portions of the anchor were determined. The lipid moiety of the anchor is an inositolphosphoceramide which contains C18:0 phytosphingosine as a long chain base, and a mixture of fatty acids with a C18:1 mono-unsaturated fatty acid as the major component. The purified GPI anchor was susceptible to digestion by a bacterial phosphatidylinositol-specific phospholipase-C enzyme. The glycan of the GPI anchor consisted of two molecular species present in the ratio 55:45, the structures of which were determined by exoglycosidase sequencing and found to be Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2 and Man alpha 1-2Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2. The glucosamine in both structures is glycosidically linked to the inositol ring of the inositolphosphoceramide. The GPI glycan structures are consistent with the conserved core structure of all characterised GPI anchors, and the structure of the D. discoideum GPI moiety has features in common with structures from yeast, protozoa and higher eukaryotes. Compositional analysis of the carbohydrate attached to threonine residues in the carboxy-terminal peptide domain is also presented. The oligosaccharides bind to wheat germ agglutinin, and contain glucosamine and fucose as the major constituents.

    AB - Prespore-specific antigen (PsA) is a cell-surface glycoprotein isolated from Dictyostelium discoideum, which is post-translationally modified by addition of carbohydrate to threonine residues of the carboxy-terminal peptide domain, and a glycosylphosphatidylinositol (GPI) anchor which attaches the glycoprotein to the cell membrane. The GPI anchor was isolated by proteolytic cleavage of the protein, and the structure of the lipid and glycan portions of the anchor were determined. The lipid moiety of the anchor is an inositolphosphoceramide which contains C18:0 phytosphingosine as a long chain base, and a mixture of fatty acids with a C18:1 mono-unsaturated fatty acid as the major component. The purified GPI anchor was susceptible to digestion by a bacterial phosphatidylinositol-specific phospholipase-C enzyme. The glycan of the GPI anchor consisted of two molecular species present in the ratio 55:45, the structures of which were determined by exoglycosidase sequencing and found to be Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2 and Man alpha 1-2Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2. The glucosamine in both structures is glycosidically linked to the inositol ring of the inositolphosphoceramide. The GPI glycan structures are consistent with the conserved core structure of all characterised GPI anchors, and the structure of the D. discoideum GPI moiety has features in common with structures from yeast, protozoa and higher eukaryotes. Compositional analysis of the carbohydrate attached to threonine residues in the carboxy-terminal peptide domain is also presented. The oligosaccharides bind to wheat germ agglutinin, and contain glucosamine and fucose as the major constituents.

    U2 - 10.1111/j.1432-1033.1993.tb18192.x

    DO - 10.1111/j.1432-1033.1993.tb18192.x

    M3 - Article

    VL - 216

    SP - 729

    EP - 737

    JO - European Journal of Biochemistry

    JF - European Journal of Biochemistry

    SN - 0014-2956

    IS - 3

    ER -