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Carbohydrate structure of Marburg virus glycoprotein
1Biochemisches Institut am Klinikum der Universität, Giessen, FRG.
Abstract:
Marburg virus was propagated in E6 cells, a cloned cell line of Vero cells, in the presence of [6-3H]glucosamine. Radiolabelled viral glycoprotein was digested with trypsin, and oligosaccharides were liberated by sequential treatment with endo-beta-N-acetylglucosaminidase H, peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F and O-glycosidase, by beta-elimination, and by alkaline hydrolysis. After fractionation by HPLC and gel filtration, glycans were characterized chromatographically, by digestion with exoglycosidases and, in part, by methylation analysis and liquid secondary ion mass spectrometry. The oligosaccharide structures thus established include oligomannosidic and hybrid-type N-glycans, as well as neutral fucosylated bi-, tri- and tetraantennary species, most of which carry an additional bisecting N-acetylglucosamine. In addition, high amounts of neutral mucin-type O-glycans with type-1 and type-2 core structures were detected. None of the glycans present in this viral glycoprotein carried sialic acid residues.
Insights
Marburg virus glycoprotein contains complex N-glycans and O-glycans, but lacks sialic acid. This detailed glycan analysis provides insights into viral structure and potential therapeutic targets.
Area of Science:
- Virology
- Glycobiology
- Biochemistry
Background:
- Marburg virus is a significant human pathogen.
- Understanding viral glycoprotein structure is crucial for developing antivirals.
- The glycosylation patterns of Marburg virus glycoproteins are not fully characterized.
Purpose of the Study:
- To characterize the N- and O-linked glycans of Marburg virus glycoprotein.
- To identify specific glycan structures and modifications.
- To investigate the presence or absence of sialic acid residues.
Main Methods:
- Propagation of Marburg virus in E6 cells with radiolabeled glucosamine.
- Enzymatic and chemical release of oligosaccharides from viral glycoprotein.
- Fractionation using High-Performance Liquid Chromatography (HPLC) and gel filtration.
- Structural characterization by exoglycosidase digestion, methylation analysis, and mass spectrometry.
Main Results:
- Oligosaccharide structures include oligomannosidic and hybrid-type N-glycans.
- Neutral fucosylated bi-, tri-, and tetraantennary N-glycans with bisecting N-acetylglucosamine were identified.
- High amounts of neutral mucin-type O-glycans with type-1 and type-2 core structures were detected.
- No sialic acid residues were found on any of the viral glycans.
Conclusions:
- Marburg virus glycoprotein exhibits complex glycosylation patterns.
- The absence of sialic acid may influence viral interactions and immune evasion.
- These findings contribute to a deeper understanding of Marburg virus biology.