Glyco

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

Abstract :

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-acetylglucosaminidase H (Endo-H) digestion, SDS-PAGE and immunoblotting
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

Output (sent_index, trigger, protein, sugar, site):

0. contains, , transferrin receptor, a high-mannose oligosaccharide, site
0. glycosylation, , transferrin receptor, -, site
1. contains, , TfR, three N-linked oligosaccharides, -
1. contains, , transferrin receptor, three N-linked oligosaccharides, -
3. present, , -, The oligosaccharide(s), site
5. contains, , -, high-mannose or Endo-H-sensitive hybrid oligosaccharides, Asn-727 site
6. Glycosylation, , -, -, Asn-727

Output(Part-Of) (sent_index, protein, site):

*Output_Site_Fusion* (sent_index, protein, sugar, site):

5. transferrin receptor, high-mannose or Endo-H-sensitive hybrid oligosaccharides, Asn-727 site
6. TfR, -, Asn-727