Analytical detection of 9(4)-O-acetylated sialoglycoproteins and gangliosides using influenza C virus
| Autor: | Catherine Dubois, Jean-Claude Manuguerra, Claude Hannoun |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2004 |
| Předmět: |
Influenzavirus C
Sialoglycoproteins Orthomyxoviridae Blotting Western Biophysics Hemagglutinin (influenza) Chick Embryo Biochemistry Virus Article chemistry.chemical_compound Epitopes Mice Gangliosides Glycophorin Animals Molecular Biology Antigens Viral Ganglioside Sheep biology Erythrocyte Membrane Esterases Antibodies Monoclonal Collodion Acetylation Cell Biology biology.organism_classification Sialic acid chemistry biology.protein Electrophoresis Polyacrylamide Gel Influenza C Virus |
| Zdroj: | Analytical Biochemistry |
| ISSN: | 1096-0309 0003-2697 |
| Popis: | The unique glycoprotein of influenza C virus, designated hemagglutinin (HEF), exhibits three functions: hemagglutination, esterase activity, and fusion factor. As the virus uses 9-O-acetylated sialic acid as a high-affinity receptor determinant for attachment to cells, its binding activity was used to reveal O-acetylated sialic acid residues after polyacrylamide gel electrophoresis and transfer onto nitrocellulose sheets of proteins and thin-layer chromatography of lipids. The specificity of the binding for O-acetylated sialoglycoconjugates was investigated. Our results showed that influenza C virus could detect the different forms of the two murine glycophorins which are known to be O-acetylated sialoglycoconjugates. The virus also bound to O-acetylated gangliosides isolated from embryonic chicken brain such as purified O-acetylated NeuAc alpha (2-8)NeuAc alpha (2-8)NeuAc alpha (2-3)Gal beta (1-4)Glc beta (1-1)ceramide (GT3). The esterase activity of the HEF protein of influenza C virus was used to unmask the sialic acid. After its deacetylation by the virus enzyme, the O-acetylated GT3 was recognized by a monoclonal antibody which binds only to the nonacetylated derivative. The results presented here show that influenza C virus is a discriminating analytical probe for identifying O-acetylated sialoglycoconjugates directly after Western blotting of proteins and thin-layer chromatography of lipids, thus providing a new analytical tool. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect