Advertisement

Release of N-glycans by Enzymatic Methods

  • Noriko Takahashi
  • Hirokazu Yagi
  • Koichi Kato

Abstract

To release oligosaccharide moiety from the original glycoprotein molecule, there are two widely used methods: the typical chemical method, hydrazinolysis; and the enzymatic method using glycoamidase A (from almond) or N-glycanase F (from Flabobacterium meningosepticum). In the past, endo-β-N-acetylglucosaminidase used to be applied, too, but the disadavantage of this enzyme is that it always leaves one of the two reducing ends N-acetylglucosamine molecules of the oligosaccharide moiety on the substrate protein molecules. On the contrary, glycoamidase A or N-glycanase F releases the whole sugar moiety completely from the substrate protein portion. Therefore, the application of the latter is the most convenient method for structural study of oligosaccharide moieties in various glycoproteins.

Keywords

Sialic Acid Enzymatic Method Sugar Moiety Amide Linkage High Performance Liquid Chroma 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Takahashi N (1977) Demonstration of a new amidase acting on glycopeptides. Biochem Biophys Res Commun 76:1194–1201PubMedCrossRefGoogle Scholar
  2. Takahashi N, Nakagawa H, Fujikawa K, Kawamura Y, Tomiya N (1995) Three-dimensional elution mapping of pyridylaminated N-linked neutral and sialyl oligosaccharides. Anal Biochem 226:139–146PubMedCrossRefGoogle Scholar
  3. Takahashi N, Cohen-Solal J, Galinha A, Fridman WH, Sautès-Fridman C, Kato K (2002) N-glycosylation profile of recombinant human soluble Fcγ receptor III. Glycobiology 12:507–515PubMedCrossRefGoogle Scholar
  4. Tomiya N, Awaya J, Kurono M, Endo S, Arata Y, Takahashi N (1988) Analyses of N-linked oligosaccharides using a two-dimensional mapping technique. Anal Biochem 171:73–90PubMedCrossRefGoogle Scholar
  5. Yagi H, Takahashi N, Yamaguchi Y, Kimura N, Uchimura K, Kannagi R, Kato K (2005) Development of structural analysis of sulfated N-glycans by multidimensional high performance liquid chromatography mapping methods. Glycobiology 15:1051–1060PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2008

Authors and Affiliations

  • Noriko Takahashi
    • 1
  • Hirokazu Yagi
    • 1
  • Koichi Kato
    • 1
  1. 1.Graduate School of Pharmaceutical SciencesNagoya City UniversityNagoyaJapan

Personalised recommendations