Characterization of the high mannose asparagine-linked oligosaccharides synthesized by Schistosoma mansoni adult male worms.
This report describes the structures of the high-mannose-type N-linked oligosaccharides in glycoproteins synthesized by Schistosoma mansoni adult male worms. Adult male schistosomes were incubated in vitro in media containing either [2-3H]mannose, [6-3H]glucosamine or [6-3H]galactose to allow metabolic radiolabeling of the oligosaccharide moieties of newly synthesized glycoproteins. Glycopeptides were prepared from the radiolabeled glycoproteins by digestion with Pronase and fractionation by chromatography on concanavalin A-Sepharose. Eleven percent of [3H]mannose incorporated into the schistosome glycopeptides was recovered in high mannose-type Asn-linked oligosaccharides which bound to the immobilized lectin. Upon treatment of [3H]mannose-labeled glycopeptide with endo-beta-N-acetylglucosaminidase H, the high mannose-type chains were released and their structures were determined by high performance liquid chromatography, methylation analysis, acetolysis and exoglycosidase digestion. The major species of high mannose-type chains synthesized by S. mansoni adult males have the composition Man7GlcNAc2, Man8GlcNac2 and Man9GlcNA2. Structural analyses indicate that these oligosaccharides are similar to high mannose-type chains synthesized by mammalian cells.
Nyame
K
K
Department of Zoology, University of Georgia, Athens 30602.
Cummings
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RD
Damian
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RT
D000818:Animals; D001216:Asparagine / Q000378:metabolism; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002854:Chromatography, Paper; D002855:Chromatography, Thin Layer; D006020:Glycopeptides / Q000032:analysis*; D006026:Glycoside Hydrolases / Q000378:metabolism; D008297:Male; D008358:Mannose / Q000378:metabolism; D008745:Methylation; D009844:Oligosaccharides / Q000032:analysis / Q000096:biosynthesis*; D012550:Schistosoma mansoni / Q000378:metabolism*
D006020:Glycopeptides; D009844:Oligosaccharides; D001216:Asparagine; D006026:Glycoside Hydrolases; D008358:Mannose