Analysis of specific interactions of synthetic glycopolypeptides carrying N-acetyllactosamine and related compounds with lectins.
Analysis of interactions of synthetic glycopolypeptides with lectins was performed with a biosensor based on surface plasmon resonance (SPR). A series of synthetic oligosaccharide-substituted poly(L-glutamic acid)s were immobilized on sensor surfaces via the gamma-carboxyl groups of their peptide moieties by the surface thiol coupling method. Artificial glycopolypeptides: an N-acetyllactosamine-substituted polymer (1), an N-acetylisolactosamine-substituted polymer (2), a (GlcNAc)3-substituted polymer (3), a (GlcNAc)2-substituted polymer (4), and a p-aminophenyl N-acetyl-beta-lactosaminide-substituted polymer (5), were used as the ligands. On analysis by SPR, surface-bound polymers 1 and 5 reacted with Erythrina cristagalli agglutinin (ECA), Lycopersicon esculentum agglutinin (LEA), Ricinus communis agglutinin-120 (RCA120), and wheat germ (Triticum vulgaris) agglutinin (WGA). Polymer 2 reacted with WGA and RCA120, but did not with ECA and LEA. The results indicate that beta-(1-->4)-linked galactosyl residues are needed for binding to ECA and LEA. Polymer 3 reacted strongly with LEA and WGA, but polymer 4 reacted strongly only with WGA. Affinity constants (KA) for surface-bound polymer 5-lectin interactions were also about 4-61 times as strong as those for surface-bound polymer 1-lectin interactions. These artificial glycopolypeptides were shown to be useful as tools and probes of carbohydrate recognition and modeling in the analysis of glycoprotein-lectin interactions.
Zeng
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Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Japan.
Murata
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Kawagishi
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Usui
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Kobayashi
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D000606:Amino Sugars / Q000032:analysis* / Q000378:metabolism; D015374:Biosensing Techniques; D002240:Carbohydrate Sequence; D006020:Glycopeptides / Q000138:chemical synthesis / Q000737:chemistry*; D007700:Kinetics; D037102:Lectins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D019887:Peanut Agglutinin / Q000378:metabolism; D037121:Plant Lectins; D013499:Surface Properties; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D000606:Amino Sugars; D006020:Glycopeptides; D037102:Lectins; D009844:Oligosaccharides; D019887:Peanut Agglutinin; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D014909:Wheat Germ Agglutinins; C045629:erythrina lectin; C034981:tomato lectin; C000458:N-acetyllactosamine