Interaction of asparagine-linked oligosaccharides with an immobilized rice (Oryza sativa) lectin column.
The carbohydrate-binding specificity of rice (Oryza sativa) lectin was investigated by testing the ability of radioactively labelled glycopeptides and oligosaccharides to bind to a rice lectin-Sepharose 4B column. Rice lectin binds asparagine-linked oligosaccharides through the core NN'-diacetylchitobiose moiety. Whereas beta 1-4-mannose enhances the binding strength only to a small extent, alpha 1-3-linked core mannose increases it considerably. A core alpha 1-6-linked mannose residue has a slightly inhibitory effect. Binding is not affected when one or both of the alpha-mannose residues are substituted with mannose at C-2, C-3 and C-6 or with N-acetylglucosamine (GlcNAc) at C-2 positions. The presence of an alpha 1-6-fucose residue attached to the asparagine-linked GlcNAc also does not affect the binding. The binding of complex biantennary glycopeptides is not altered by the presence of one or two galactose residues in the non-reducing terminus, but the presence of one or two sialic acid residues decreases the binding capacity. A bisecting beta 1-4-linked GlcNAc attached to beta-linked mannose residue enhances the binding of sialo, asialo and asialoagalacto complex biantennary-type glycopeptides. Bisected hybrid-type glycopeptides bind very tightly to a rice lectin-Sepharose 4B column: Substitution of alpha 1-3-mannose residue at C-2 and C-4 with GlcNAc completely inhibits the binding of both bisected and non-bisected complex asparagine-linked glycopeptides. O-Glycosidically linked oligosaccharides containing GlcNAc bind very weakly to a rice lectin column. The applicability of immobilized rice lectin columns in the fractionation of asparagine-linked oligosaccharides is discussed.
Poola
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Department of Biochemistry and Molecular Biology, SUNY Health Science Center, Syracuse 13210.
Narasimhan
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D001216:Asparagine; D002846:Chromatography, Affinity; D006020:Glycopeptides; D037102:Lectins; D046911:Macromolecular Substances; D009844:Oligosaccharides / Q000302:isolation & purification*; D037121:Plant Lectins; D013329:Structure-Activity Relationship
D006020:Glycopeptides; D037102:Lectins; D046911:Macromolecular Substances; D009844:Oligosaccharides; D037121:Plant Lectins; C040599:rice lectin; D001216:Asparagine