Structural analysis and localization of the carbohydrate moieties of a soluble human interferon gamma receptor produced in baculovirus-infected insect cells

Protein Sci. 1994 Jan;3(1):30-8. doi: 10.1002/pro.5560030105.


A soluble form of the human interferon gamma receptor that is required for the identification of interferon gamma antagonists was expressed in baculovirus-infected insect cells. The protein carried N-linked carbohydrate and showed a heterogeneity on denaturing polyacrylamide gels. We investigated the utilization of the potential sites for N-linked glycosylation and the structure of the carbohydrate moieties of this soluble receptor. Amino acid sequence analysis and ion spray mass spectrometry revealed that of the five potential sites for N-linked glycosylation, Asn17 and Asn69 were always utilized, whereas Asn62 and Asn162 were utilized in approximately one-third of the protein population. Asn223 was never found to be glycosylated. The soluble receptor was treated with N-glycosidase F and the oligosaccharides released were analyzed by matrix-assisted laser desorption mass spectrometry, which showed that the protein carried six types of short carbohydrate chains. The predominant species was a hexasaccharide of molecular mass 1,039, containing a fucose subunit linked to the proximal N-acetylglucosamine residue: [formula: see text]

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Baculoviridae / genetics*
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Carbohydrates / analysis
  • Carbohydrates / chemistry*
  • Chromatography, High Pressure Liquid
  • Genetic Vectors
  • Glycosylation
  • Humans
  • Molecular Sequence Data
  • Moths*
  • Receptors, Interferon / chemistry*
  • Receptors, Interferon / genetics
  • Recombinant Proteins / chemistry


  • Carbohydrates
  • Receptors, Interferon
  • Recombinant Proteins