Determination of lectin-sugar binding constants by microequilibrium dialysis coupled with high performance liquid chromatography.
We used high performance liquid chromatography to determine the concentrations of free ligands after equilibrium dialysis to examine lectin-sugar interactions. The binding of p-nitrophenyl 1-thio-alpha-mannoside, p-nitrophenyl N-acetyl-1-thio-beta-glucosaminide, and the pyridylamino derivative of Man6GlcNAc2 to concanavalin A and Triticum vulgaris lectin was examined. The binding constant, Ka, and the concentration of total binding sites, [L]t, were calculated from the trace amounts of sugars and lectins using the equation, [S]/[LS] = 1/Ka[L]t + [S]/[L]t, where [S] is the concentration of free ligand and [LS] the concentration of bound ligand.
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Department of Chemistry, Osaka University College of Science.
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D002241:Carbohydrates / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003956:Dialysis / Q000295:instrumentation / Q000379:methods; D007700:Kinetics; D037102:Lectins / Q000737:chemistry*; D008024:Ligands; D008433:Mathematics; D008962:Models, Theoretical; D011485:Protein Binding; D013997:Time Factors
D002241:Carbohydrates; D037102:Lectins; D008024:Ligands