Carbohydrates of influenza virus. Structure of the oligosaccharides linked to asparagines 406 and 478 in the hemagglutinin of fowl plague virus,strain dutch |
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Authors: | Rudolf Geyer Silvia Diabaté Hildegard Geyer Hans-Dieter Klenk Heiner Niemann Stephan Stirm |
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Affiliation: | (1) Biochemisches Institut am Klinikum der Universität, Friedrichstrasse 24, D-6300 Giessen, Federal Republic of Germany;(2) Institut für Medizinische Virologie am Klinikum der Universität, Frankfurter Strasse 107, D-6300 Giessen, Federal Republic of Germany |
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Abstract: | Fowl plague virus, strain Dutch, was metabolically labeled withd-[2-3H]mannose, or withd-[6-3H]glucosamine, and the small subunit (HA2; 0.8 mg in total) of the viral hemagglutinin was isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis. After proteolytic digestion, the radioactive oligosaccharides were sequentially liberated from the glycopeptides by treatment with different endo- -N-acetylglucosaminidases and with peptide:N-glycosidase or, finally, by hydrazinolysis. In this manner, four groups of glycans could be obtained by consecutive gel filtrations and were subfractionated by HPLC. The structures of the individual oligosaccharides were analyzed by micromethylation, by acetolysis or by digestion with exoglycosidases. The major species amongst the high mannose glycans at Ans-406 of the viral glycopolypeptide were found to be Man 1-2Man 1-3(Man 1-2Man 1-6)Man 1-6(Man 1-2Man 1-2Man 1-3)Man 1-4GlcNac 1-4GlcNAc and Man 1-3(Man 1-2Man 1-6)Man 1-6(Man 1-2Man 1-2Man 1-3)Man 1-4GlcNAc 1-4GlcNAc, while the complex glycans at Asn-478 are predominantly GlcNAc 1-2Man 1-3(GlcNAc 1-2Man 1-6)Man 1-4GlcNAc 1-4GlcNAc (lacking, in part, one of the outerN-acetylglucosamine residues) and GlcNAc 1-2Man 1-3(Gal 1-4GlcNAc 1-2Man 1-6)Man 1-4GlcNAc 1-4GlcNAc.Abbreviation BSA bovine serum albumin - endo D (F,H) endo- -N-acetyl-d-glucosaminidase D (F,H) - HA hemagglutinin (HA1, large subunit of HA - HA2 small subunit - FPV fowl plague virus - PNGase F peptide:N-glycosidase F - SDS sodium dodecylsulfate |
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Keywords: | influenza virus hemagglutinin glycoprotein glycans oligosaccharide structure |
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