Analysis of the neutral glycan fractions of glycosyl-phosphatidylinositols by thin-layer chromatography

P. Schneider, J. E. Ralton, M. J. McConville, M. A. J. Ferguson

    Research output: Contribution to journalArticle

    27 Citations (Scopus)

    Abstract

    The radiolabeled neutral glycan fractions of both glycosyl-phosphatidylinositol (GPI) protein anchors and related glycolipids were analyzed by thin-layer chromatography on silica gel 60, using butanol:ethanol:water (4:3:3, v/v) or a combination of 1-propanol:acetone:water (9:6:5, v/v and 5:4:1, v/v) as solvents. Dextran acid hydrolysates were used as standards, and oligomers up to 11 glucose units could be resolved. A comparison of 18 GPI-glycan standards revealed that their migration was dependent mainly on the size of the oligosaccharide. Isomers were generally not resolved, with the exception of Mana1-6Mana1-4AHM and Mana1-3Mana1-4AHM. Structures containing galactofuranose or GalNAc were well resolved from structures containing only Galp and/or Manp. The utility of this method for the microsequencing of radiolabeled neutral glycans derived from two GPI glycolipids, using exoglycosidases and chemical treatments, is demonstrated. This method is a simple and useful complement to the existing chromatographic techniques.
    Original languageEnglish
    Pages (from-to)106-112
    Number of pages7
    JournalAnalytical Biochemistry
    Volume210
    Issue number1
    DOIs
    Publication statusPublished - Apr 1993

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    Thin layer chromatography
    Glycosylphosphatidylinositols
    Thin Layer Chromatography
    Polysaccharides
    Glycolipids
    1-Propanol
    Butanols
    Water
    Glycoside Hydrolases
    Silica Gel
    Acetone
    Dextrans
    Anchors
    Oligosaccharides
    Oligomers
    Isomers
    Ethanol
    Glucose
    Acids
    Proteins

    Cite this

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    title = "Analysis of the neutral glycan fractions of glycosyl-phosphatidylinositols by thin-layer chromatography",
    abstract = "The radiolabeled neutral glycan fractions of both glycosyl-phosphatidylinositol (GPI) protein anchors and related glycolipids were analyzed by thin-layer chromatography on silica gel 60, using butanol:ethanol:water (4:3:3, v/v) or a combination of 1-propanol:acetone:water (9:6:5, v/v and 5:4:1, v/v) as solvents. Dextran acid hydrolysates were used as standards, and oligomers up to 11 glucose units could be resolved. A comparison of 18 GPI-glycan standards revealed that their migration was dependent mainly on the size of the oligosaccharide. Isomers were generally not resolved, with the exception of Mana1-6Mana1-4AHM and Mana1-3Mana1-4AHM. Structures containing galactofuranose or GalNAc were well resolved from structures containing only Galp and/or Manp. The utility of this method for the microsequencing of radiolabeled neutral glycans derived from two GPI glycolipids, using exoglycosidases and chemical treatments, is demonstrated. This method is a simple and useful complement to the existing chromatographic techniques.",
    author = "P. Schneider and Ralton, {J. E.} and McConville, {M. J.} and Ferguson, {M. A. J.}",
    year = "1993",
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    doi = "10.1006/abio.1993.1158",
    language = "English",
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    Analysis of the neutral glycan fractions of glycosyl-phosphatidylinositols by thin-layer chromatography. / Schneider, P.; Ralton, J. E.; McConville, M. J.; Ferguson, M. A. J.

    In: Analytical Biochemistry, Vol. 210, No. 1, 04.1993, p. 106-112.

    Research output: Contribution to journalArticle

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    AU - Schneider, P.

    AU - Ralton, J. E.

    AU - McConville, M. J.

    AU - Ferguson, M. A. J.

    PY - 1993/4

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    N2 - The radiolabeled neutral glycan fractions of both glycosyl-phosphatidylinositol (GPI) protein anchors and related glycolipids were analyzed by thin-layer chromatography on silica gel 60, using butanol:ethanol:water (4:3:3, v/v) or a combination of 1-propanol:acetone:water (9:6:5, v/v and 5:4:1, v/v) as solvents. Dextran acid hydrolysates were used as standards, and oligomers up to 11 glucose units could be resolved. A comparison of 18 GPI-glycan standards revealed that their migration was dependent mainly on the size of the oligosaccharide. Isomers were generally not resolved, with the exception of Mana1-6Mana1-4AHM and Mana1-3Mana1-4AHM. Structures containing galactofuranose or GalNAc were well resolved from structures containing only Galp and/or Manp. The utility of this method for the microsequencing of radiolabeled neutral glycans derived from two GPI glycolipids, using exoglycosidases and chemical treatments, is demonstrated. This method is a simple and useful complement to the existing chromatographic techniques.

    AB - The radiolabeled neutral glycan fractions of both glycosyl-phosphatidylinositol (GPI) protein anchors and related glycolipids were analyzed by thin-layer chromatography on silica gel 60, using butanol:ethanol:water (4:3:3, v/v) or a combination of 1-propanol:acetone:water (9:6:5, v/v and 5:4:1, v/v) as solvents. Dextran acid hydrolysates were used as standards, and oligomers up to 11 glucose units could be resolved. A comparison of 18 GPI-glycan standards revealed that their migration was dependent mainly on the size of the oligosaccharide. Isomers were generally not resolved, with the exception of Mana1-6Mana1-4AHM and Mana1-3Mana1-4AHM. Structures containing galactofuranose or GalNAc were well resolved from structures containing only Galp and/or Manp. The utility of this method for the microsequencing of radiolabeled neutral glycans derived from two GPI glycolipids, using exoglycosidases and chemical treatments, is demonstrated. This method is a simple and useful complement to the existing chromatographic techniques.

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