Characterisation of the asparagine‐linked oligosaccharides from Trypanosoma brucei type‐I variant surface glycoproteins

• Published in: European journal of biochemistry Vol. 187; no. 3; pp. 657 - 663
• Main Authors: ZAMZE, Susanne E., WOOTEN, E. Wrenn, ASHFORD, David A., FERGUSON, Michael A. J., DWEK, Raymond A., RADEMACHER, Thomas W.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 14.02.1990; Blackwell
• Subjects: Acetone; Analytical, structural and metabolic biochemistry; Animals; Asparagine - analysis; Binding Sites; Biological and medical sciences; Blood - parasitology; Borohydrides; Carbohydrate Sequence; Carbohydrates; Chromatography - methods; Cloning, Molecular; Electrophoresis, Paper; Fundamental and applied biological sciences. Psychology; Glycoside Hydrolases; Glycosylation; Magnetic Resonance Spectroscopy; Membrane Glycoproteins - analysis; Membrane Glycoproteins - isolation & purification; Methylation; Molecular Sequence Data; Oligosaccharides - analysis; Other biological molecules; Peptide Fragments - analysis; Rats; Trypanosoma brucei; Trypanosoma brucei brucei - analysis; Trypanosoma brucei brucei - genetics; Protozoa; Oligosaccharide; Rhizoflagellata; Acetolysis; Enzymatic digestion; Primary structure; Glycosylation; NMR spectrometry; Methylation; Trypanosoma brucei; Site of action; Gel permeation chromatography
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1990.tb15350.x

The complete primary structures of the Asn‐linked oligosaccharides from the conserved glycosylation site of the type‐I variant surface glycoproteins of Trypanosoma brucei MITat 1.4 and MITat 1.6 were determined using a combination of exoglycosidase digestions, permethylation analysis, acetolysis and 1H NMR. Both variants contained almost exclusively oligomannose‐type oligosaccharides, identical in structure to those of mammalian glycoproteins. The oligosaccharides ranged in size from (Man)9(GlcNAc)2 to (Man)5(GlcNAc)2. The relative abundance of each component was similar in both variants. The major components were (Man)8(GlcNAc)2 and (Man)7(GlcNAc)2 with slightly less (Man)9(GlcNAc)2 and (Man)6(GlcNAc)2 and much less (Man)5(GlcNAc)2. Both variants also contained the same structural isomers. The close similarity of the oligomannose series indicates identical processing at the conserved site in both variants.