Use of hydrazine to release in intact and unreduced form both N- and O-linked oligosaccharides from glycoproteins

Biochemistry. 1993 Jan 19;32(2):679-93. doi: 10.1021/bi00053a037.


The use of hydrazine to release unreduced N- and O-linked oligosaccharides from glycoproteins has been investigated using several "standard" glycoproteins of previously defined glycosylation. It is shown that hydrazinolysis can be used to release intact N- and O-linked oligosaccharides in an unreduced form. The release of O-linked oligosaccharides occurs with a lower temperature dependence than the release of N-linked oligosaccharides, and the kinetic parameters governing release of oligosaccharides from these standard glycoproteins have been determined. These parameters allow a definition of reaction conditions under which anhydrous hydrazinolysis can be used to selectively release O-linked oligosaccharides (60 degrees C, 5 h) or release both N- and O-linked oligosaccharides (95 degrees C, 4 h) in high yield (> 85%) from all glycoproteins investigated (n = 11). Under these reaction conditions, the recovered N- and O-linked oligosaccharides are structurally intact (as judged by 600-MHz 1H-NMR, laser-desorption mass spectrometry, HPAEC-PAD, gel filtration, and glycosidase digestion), with the possible exception of certain N- and O-acyl substituents of sialic acid. This use of mild hydrazinolysis therefore allows both the simultaneous and sequential chemical release from glycoproteins of O- and N-linked oligosaccharides in their intact unreduced form.

MeSH terms

  • Animals
  • Carbohydrate Sequence
  • Cattle
  • Chickens
  • Glycoproteins / chemistry*
  • Humans
  • Hydrazines / chemistry*
  • Hydrolysis
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Mass Spectrometry
  • Molecular Sequence Data
  • Oligosaccharides / chemistry*
  • Oxidation-Reduction
  • Peptides / chemistry
  • Swine
  • Temperature


  • Glycoproteins
  • Hydrazines
  • Oligosaccharides
  • Peptides
  • hydrazine