Glycoconjugate Journal

, Volume 14, Issue 2, pp 225–229 | Cite as

Alteration of N-linked oligosaccharide structures of human chorionic gonadotropin β-subunit by disruption of disulfide bonds

  • Takayuki Moriwaki
  • Nobuhiko Suganuma
  • Madoka Furuhashi
  • Fumitaka Kikkawa
  • Yutaka Tomoda
  • Irving Boime
  • Munehiro Nakata
  • Tsuguo Mizuochi
Article

Abstract

The human chorionic gonadotropin β-subunit (hCGβ) is a glycoprotein in which 12 cysteine residues pair to form six intramolecular disulfide bonds. In order to elucidate the effect of each disulfide bond on glycosylation of the molecule, we analysed structures of asparagine-linked oligosaccharides of various recombinant hCGβ produced in Chinese hamster ovary (CHO) cells: wild-type hCGβ (βWT) and mutants in which any one of the six intramolecular disulfide bonds had been disrupted by site-directed mutagenesis. SDS-PAGE analysis of βWT and these mutants before and after digestion with endoglycosidase F and H revealed structural changes in the oligosaccharide moieties of some mutants. In addition, structural analysis of oligosaccharides obtained from metabolically labeled βWT and a mutant showed that the mutant contained additional high mannose type oligosaccharides. These results suggest that elimination of a specific disulfide bond, resulting in a change in the protein conformation, disturbs the normal assembly of the mature complex type oligosaccharides in the hCGβ molecule. Abbreviations: hCGβ, human chorionic gonadotropin β-subunit; βWT, wild type hCGβ; CHO, Chinese hamster ovary; Endo-H, endoglycosidase H; Endo-F, endoglycosidase F

human chorionic gonadotropin hCGβ-subunit disulfide bond N-linked oligosaccharide processing 

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Copyright information

© Chapman and Hall 1997

Authors and Affiliations

  • Takayuki Moriwaki
    • 1
  • Nobuhiko Suganuma
    • 1
  • Madoka Furuhashi
    • 1
  • Fumitaka Kikkawa
    • 1
  • Yutaka Tomoda
    • 1
  • Irving Boime
    • 2
  • Munehiro Nakata
    • 3
  • Tsuguo Mizuochi
    • 3
  1. 1.Department of Obstetrics and GynecologyNagoya University School of MedicineNagoya AichiJapan
  2. 2.Department of Molecular Biology and PharmacologyWashington University School of MedicineSt LouisUSA
  3. 3.Laboratory of Biomedical Chemistry, Department of Applied ChemistryTokai University HiratsukaKanagawaJapan

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