N-glycan structures of a recombinant mouse soluble Fcgamma receptor II

Glycoconj J. 1998 Sep;15(9):905-14. doi: 10.1023/a:1006915200989.

Abstract

N-glycans of a recombinant mouse soluble Fcgamma receptor II (sFcgammaRII) expressed in baby hamster kidney cells were released from glycopeptides by digestion with glycoamidase A (from sweet almond), and the reducing ends of the oligosaccharides were reductively aminated with 2-aminopyridine. The derivatized N-glycans were separated and structurally identified by a three-dimensional high-performance liquid chromatography (HPLC) mapping technique on three kinds of HPLC columns [Takahashi, et al. (1995) Anal. Biochem. 226:139-46]. Eighteen different major N-glycan structures were identified, of which six were neutral (45%), five mono-sialyl (49%), one di-sialyl (4.6%), five tri-sialyl (1.1%), and one tetra-sialyl (0.3%). All N-glycan structures determined were complex type with fucosylation at the N-acetylglucosamine residue of the reducing end, and N-acetylneuraminic acid, when present, was alpha-(2,3)-linked. The existence of a unique structure containing both N-acetylgalactosamine and alpha-(2,3)-N-acetylneuraminic acid residues at the reducing ends, as below, was confirmed by MALDI-TOF mass spectrometry. Carbohydrate sequence [see text]

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Glycoproteins / chemistry
  • Mice
  • Molecular Sequence Data
  • Oligosaccharides / chemistry
  • Polysaccharides / chemistry*
  • Receptors, IgG / chemistry*
  • Receptors, IgG / genetics
  • Recombinant Proteins / chemistry
  • Solubility
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Glycoproteins
  • Oligosaccharides
  • Polysaccharides
  • Receptors, IgG
  • Recombinant Proteins