The critical glycosylation site of human transferrin receptor contains a high-mannose oligosaccharide

Gary R. Hayes, Anthony Williams, Catherine E. Costello, Caroline A. Enns, John J. Lucas

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


The human transferrin receptor (TfR) contains three N-linked oligosaccharides and glycosylation is required for the proper folding and function of the molecule. Earlier studies demonstrated that the oligosaccharide at Asn-727 is vital for the production of fully active TfR. The oligosaccharide(s) present at this site have been analysed using a combination of site-directed mutagenesis and chemical analysis. Wild-type TfR and mutants containing only the Asn-727 site or missing all three sites were transfected into mouse 3T3 cells and receptors were analysed by endo-N-acetylglucosam-inidase H (Endo-H) digestion, SDS-PAGE and immuno-blotting. These studies suggested that the Asn-727 site contains high-mannose or Endo-H-sensitive hybrid oligosaccharides. Glycosylation of Asn-727 found in the TfR purified from human placentae was analysed by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) and mass spectrometry following tryptic digestion, peptide purification via reverse-phase high-performance liquid chromatography (RP-HPLC) and peptide sequencing. HPAE-PAD showed the presence of a series of high-mannose oligosaccharides. Mass spectrometry confirmed these observations, but also showed the presence of an 80 Da anionic moiety on a fraction of the oligosaccharides.

Original languageEnglish (US)
Pages (from-to)227-232
Number of pages6
Issue number2
StatePublished - Mar 1995


  • Characterization
  • Glycosylation site
  • Human
  • Oligosacharides
  • Transferrin receptor

ASJC Scopus subject areas

  • Biochemistry


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