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Strategy for the investigation of O-linked oligosaccharides from mucins based on the separation into neutral, sialic acid- and sulfate-containing species.

Journal article
Authors Niclas G. Karlsson
Hasse Karlsson
Gunnar C. Hansson
Published in Glycoconjugate journal
Volume 12
Issue 1
Pages 69-76
ISSN 0282-0080
Publication year 1995
Published at Institute of Medical Biochemistry
Pages 69-76
Language en
Links www.ncbi.nlm.nih.gov/entrez/query.f...
Keywords Animals, Carbohydrate Sequence, Chemical Fractionation, Chromatography, Ion Exchange, methods, Esterification, Gas Chromatography-Mass Spectrometry, Glycopeptides, chemistry, metabolism, Intestine, Small, chemistry, Methylation, Molecular Sequence Data, Monosaccharides, chemistry, Mucins, chemistry, metabolism, Oligosaccharides, analysis, chemistry, metabolism, Sialic Acids, chemistry, Solvents, Sugar Alcohols, analysis, chemistry, metabolism, Sulfates, chemistry, Swine
Subject categories Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

Abstract

A method for the separation of O-linked oligosaccharides into neutral, sialic acid-containing and sulfated species was applied to oligosaccharides released by alkaline borohydride from mucin glycopeptides from porcine small intestine. The released mixture of reduced oligosaccharides was applied to an anion exchange column, and the neutral oligosaccharides were collected as the unretarded fraction. A mixture of dimethyl sulfoxide and iodomethane was passed through the column to convert the sialic acid-containing oligosaccharides into methyl esters that were eluted and converted to methyl amides by methyl amine. Finally the sulfated oligosaccharide fraction was eluted with salt. The neutral and the derivatized sialic acid-containing oligosaccharides were analysed by gas chromatography-mass spectrometry after permethylation and the sulfated oligosaccharide fraction was analysed by high performance anion exchange chromatography.

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