Comparison of the carbohydrate moieties of recombinant soluble Fc epsilon receptor (sFc epsilon RII/sCD23) expressed in Saccharomyces cerevisiae and Chinese hamster ovary cells. Different O-glycosylation sites are used by yeast and mammalian cells

Glycoconj J. 1992 Aug;9(4):209-16. doi: 10.1007/BF00731167.

Abstract

Recombinant human soluble low affinity receptor for the Fc portion of IgE (sFc epsilon RII/sCD23) was produced in Saccharomyces cerevisiae or Chinese hamster ovary cells and subjected to carbohydrate analysis. Applied methods included analytical SDS-PAGE, reversed phase HPLC, methylation analysis and sequential degradation with exoglycosidases. The results revealed that sFc epsilon RII derived from Chinese hamster ovary cells is glycosylated exclusively at Ser-147, containing mainly the trisaccharide Sia(alpha 2-3)Gal(beta 1-3)GalNAc, whereas the yeast derived glycoprotein was glycosylated at Ser-167 and contained only alpha-mannosyl residues. It is shown here for the first time that different amino acids of a given protein can be O-glycosylated when expressed in yeast or Chinese hamster ovary cells.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CHO Cells
  • Carbohydrate Sequence
  • Carbohydrates / analysis*
  • Cloning, Molecular
  • Cricetinae
  • Electrophoresis, Polyacrylamide Gel
  • Glycosylation
  • Humans
  • Mass Spectrometry
  • Molecular Sequence Data
  • Peptide Mapping
  • Receptors, IgE / chemistry*
  • Receptors, IgE / genetics
  • Receptors, IgE / isolation & purification
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Saccharomyces cerevisiae

Substances

  • Carbohydrates
  • Receptors, IgE
  • Recombinant Proteins