Carbohydrate masking of an antigenic epitope of influenza virus haemagglutinin independent of oligosaccharide size.
Comparison of the haemagglutinins (HA) of the pathogenic avian influenza viruses A/FPV/Dutch/27 (H7N7) and A/FPV/Rostock/34 (H7N1) revealed 94.7% nucleotide and 93.8% amino acid sequence homologies. Six of the seven N-glycosidic oligosaccharides of the Rostock HA are at the same positions as the six carbohydrates of the Dutch strain. The additional oligosaccharide side chain of the Rostock strain, which is of the complex type, is attached to asparagine149 in antigenic epitope B. The accessibility of this antigenic epitope has been analysed by using rabbit antisera raised against synthetic peptides comprising amino acids 143-162. The carbohydrates of the HA of the Rostock strain have been modified (i) to truncated cores by expression in insect cells using a baculovirus vector, (ii) to oligomannosidic side chains by growth in the presence of the trimming inhibitor methyldeoxynojirimycin and (iii) to a single N-acetylglucosamine residue by removal of the oligomannosidic sugar with endo-beta-N-acetylglucosaminidase H. Neither the authentic nor the modified oligosaccharides allowed antibody binding, as indicated by enzyme-linked immunosorbent assay (ELISA) and Western blot analyses. Reactivity was observed, however, after complete removal of the carbohydrate from HA of the Rostock strain by digestion with peptide-N-glycosidase F. HA of the Dutch strain was reactive without prior peptide-N-glycosidase F treatment. These results demonstrate that a single N-acetyl-glucosamine at asparagine149 is sufficient to prevent recognition of the peptide epitope.
Munk
K
K
Institut für Virologie, Philipps-Universität Marburg, FRG.
Pritzer
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Kretzschmar
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Gutte
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B
Garten
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Klenk
H D
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D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000737:chemistry; D000939:Epitopes / Q000737:chemistry; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000276:immunology; D009980:Influenza A virus / Q000737:chemistry* / Q000276:immunology*; D008958:Models, Molecular; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000276:immunology*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D000956:Antigens, Viral; D000939:Epitopes; D006389:Hemagglutinins, Viral; D009844:Oligosaccharides