1H NMR characterization of a hen ovalbumin tyrosinamide N-linked oligosaccharide library

Arch Biochem Biophys. 1995 Apr 20;318(2):465-75. doi: 10.1006/abbi.1995.1255.


A library of 15 N-linked oligosaccharide structures was prepared from ovalbumin and characterized using high-field NMR and mass spectrometry. The oligosaccharides were enzymatically released from ovalbumin glycopeptides, and the reducing ends were reacted with ammonium bicarbonate to form oligosaccharide-glycosylamines. These reacted with Boc-tyrosine-N-hydroxysuccinimide ester, resulting in a mixture of tyrosinamide-oligosaccharides. The Boc group was removed to expose an amine terminus which enhanced the resolution of tyrosinamide-oligosaccharides when chromatographed on reverse-phase HPLC. Ten major and five minor oligosaccharides were purified on a micromole scale and characterized using 1H NMR and FAB-MS. The structures include high-mannose, hybrid, and complex oligosaccharides possessing from two to five antenna, providing the most complete definition of ovalbumin N-linked oligosaccharides to date. The resulting library is well suited to biological studies due to the presence of a single terminal tyrosine residue on each oligosaccharide that allows radioiodination or the attachment of additional probes to these glycoconjugates prior to biological studies.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Chickens
  • Chromatography, High Pressure Liquid
  • Databases, Factual
  • Female
  • Glycopeptides / chemistry
  • Glycopeptides / isolation & purification
  • Hydrogen
  • Magnetic Resonance Spectroscopy / methods
  • Molecular Sequence Data
  • Oligosaccharides / chemistry*
  • Oligosaccharides / isolation & purification
  • Ovalbumin / chemistry*
  • Spectrometry, Mass, Fast Atom Bombardment
  • Trypsin
  • Tyrosine / analogs & derivatives*


  • Glycopeptides
  • Oligosaccharides
  • tyrosinamide
  • Tyrosine
  • Hydrogen
  • Ovalbumin
  • Trypsin