Production and HPLC-Based Disaccharide Analysis of Xyloside-Primed Glycosaminoglycans

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Abstract

Although glycosaminoglycans (GAGs) are known to be involved in a variety of physiological and pathological processes, knowledge about their expression by cells or tissues, the GAGome, is limited. Xylosides can be used to induce the formation of GAGs without the presence of a proteoglycan core protein. The administration of xylosides to living cells tends to result in a considerable amplification in GAG production, and the xylosides can, therefore, be used as analytical tools to study the GAG produced by a certain cell type. One of the most common ways to analyze the GAGs structurally is by disaccharide analysis, which involves depolymerization of the GAGs into disaccharides, fluorescent labeling of the disaccharides with 2-aminoacridone, and quantification using high-pressure liquid chromatography (HPLC). Here, we describe the procedure of producing xyloside-primed GAGs and how to study them structurally by disaccharide analysis.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages173-182
Number of pages10
DOIs
Publication statusPublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2303
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Subject classification (UKÄ)

  • Cell and Molecular Biology

Free keywords

  • Chondroitin sulfate
  • Chondroitinase
  • Dermatan sulfate
  • Glycosaminoglycan
  • Heparan sulfate
  • Heparinase
  • High-pressure liquid chromatography (HPLC)
  • Proteoglycan
  • Structural analysis
  • Xyloside

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