title: 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.
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.
authors.affiliation: Ernst Boehringer Institut für Arzneimittelforschung, Bender & Co Ges mbH, Vienna, Austria.
authors.forename: I
authors.initials: I
authors.lastname: Kalsner
authors.forename: F J
authors.initials: FJ
authors.lastname: Schneider
authors.forename: R
authors.initials: R
authors.lastname: Geyer
authors.forename: H
authors.initials: H
authors.lastname: Ahorn
authors.forename: I
authors.initials: I
authors.lastname: Maurer-Fogy
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis*; D003001:Cloning, Molecular; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D017455:Receptors, IgE / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae
chemical_list: D002241:Carbohydrates; D017455:Receptors, IgE; D011994:Recombinant Proteins