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Related Experiment Videos

Analyses of N-linked oligosaccharides using a two-dimensional mapping technique.

N Tomiya1, J Awaya, M Kurono

  • 1Mie Research Laboratory, Sanwa Kagaku Kenkyusho Co. Ltd, Mie, Japan.

Analytical Biochemistry
|May 15, 1988
PubMed
Summary

We developed a novel two-dimensional sugar map for analyzing N-linked oligosaccharide structures. This method simplifies characterization and identified two new structures in hen ovalbumin.

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Area of Science:

  • Glycobiology
  • Analytical Chemistry
  • Biochemistry

Background:

  • N-linked oligosaccharides play crucial roles in biological processes.
  • Accurate structural analysis of these complex carbohydrates is essential for understanding their functions.
  • Existing methods for oligosaccharide structural determination can be complex and time-consuming.

Purpose of the Study:

  • To develop a simple, reproducible, and sensitive method for N-linked oligosaccharide structural analysis.
  • To create a two-dimensional sugar map for characterizing unknown oligosaccharide structures.
  • To identify novel oligosaccharide structures in biological samples.

Main Methods:

  • Preparation of oligosaccharides from glycopeptides using N-oligosaccharide glycopeptidase.

Related Experiment Videos

  • Fluorescent derivatization of oligosaccharide reducing ends with 2-amino-pyridine.
  • Separation and size analysis of oligosaccharide derivatives using high-performance liquid chromatography (HPLC) on ODS-silica and amide-silica columns.
  • Structural characterization via sequential exoglycosidase digestion and plotting on a two-dimensional sugar map.

    • Main Results:

    • A comprehensive database of 113 standard oligosaccharides was established for the sugar map.
    • The two-dimensional sugar map method allows for characterization of unknown oligosaccharide structures based on their position.
    • Application to hen ovalbumin identified two previously unreported unusual oligosaccharides.

    Conclusions:

    • The two-dimensional sugar map method provides a robust platform for N-linked oligosaccharide structural analysis.
    • This technique simplifies and enhances the sensitivity of oligosaccharide structure determination.
    • The discovery of novel oligosaccharides in hen ovalbumin highlights the method's utility in uncovering new biological structures.