PMID: 7780197

 

    Legend: Sugar

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

Abstract :
  1. The human transferrin receptor ( TfR ) contains three N-linked oligosaccharides and glycosylation is required for the proper folding and function of the molecule
  2. Earlier studies demonstrated that the oligosaccharide at Asn-727 is vital for the production of fully active TfR
  3. The oligosaccharide(s) present at this site have been analysed using a combination of site-directed mutagenesis and chemical analysis
  4. 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-acetylglucosaminidase H (Endo-H) digestion, SDS-PAGE and immunoblotting
  5. These studies suggested that the Asn-727 site contains high-mannose or Endo-H-sensitive hybrid oligosaccharides.
  6. 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
  7. HPAE-PAD showed the presence of a series of high-mannose oligosaccharides.
  8. Mass spectrometry confirmed these observations, but also showed the presence of an 80 Da anionic moiety on a fraction of the oligosaccharides