Characterization of oligosaccharide structures on a chimeric respiratory syncytial virus protein expressed in insect cell line Sf9.
The oligosaccharide structures added to a chimeric protein (FG) composed of the extracellular domains of respiratory syncytial virus F and G proteins, expressed in the insect cell line Sf9, were investigated. Cells were labeled in vivo with [3H]glucosamine and infected with a recombinant baculovirus containing the FG gene. The secreted chimeric protein was isolated by immunoprecipitation and subjected to oligosaccharide analysis. The FG protein contains two types of O-linked oligosaccharides: GalNAc and Gal beta 1-3GalNAc constituting 17 and 66% of the total number of structures, respectively. Only one type of N-linked oligosaccharide, constituting the remaining 17% of the structures on FG, was detected: a trimannosyl core structure with a fucose residue linked alpha 1-6 to the asparagine-linked N-acetylglucosamine.
Wathen
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Upjohn Company, Kalamazoo, Michigan 49001.
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Elhammer
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D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D016367:Baculoviridae / Q000235:genetics; D002240:Carbohydrate Sequence; D002460:Cell Line; D002678:Chimera; D005814:Genes, Viral; D005944:Glucosamine / Q000378:metabolism; D015707:HN Protein; D006801:Humans; D007313:Insecta; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification*; D012136:Respiratory Syncytial Viruses / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics; D014764:Viral Proteins
D000956:Antigens, Viral; D015707:HN Protein; D009844:Oligosaccharides; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C116204:attachment protein G; D005944:Glucosamine