The affinities of the carbohydrate-binding protein concanavalin A (Con A) for mono- and multivalent
ligands were measured by surface plasmon resonance (SPR) detection. Assessing protein−carbohydrate affinities
is typically difficult due to weak affinities observed and the complications that arise from the importance of
multivalency in these interactions. We describe a convenient method to rapidly evaluate the inhibitory constants
for a panel of different ligands, both monovalent and multivalent, for low-affinity receptors, such as the
carbohydrate-binding protein Con A. A nonnatural, mannose-substituted glycolipid was synthesized, and self-assembled monolayers of varying carbohydrate density were generated. The synthetic surfaces bind Con A.
Competition experiments that employed monovalent ligands in solution yielded K
i values similar to equilibrium
binding constants obtained in titration microcalorimetry experiments. In addition, this assay could be used to
examine various polymeric ligands of defined lengths, generated by ring-opening metathesis polymerization
(ROMP). This study demonstrates the utility of this method for rapidly screening ligands that engage in low
affinity interactions with their target receptors. Our results emphasize that those molecules that can
simultaneously occupy two or more saccharide binding sites within a lectin oligomer are effective inhibitors
of protein−carbohydrate interactions.
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