Infoscience

Journal article

Ligand binding to nicotinic acetylcholine receptor investigated by surface plasmon resonance

Ligand binding to the nicotinic acetylcholine receptor is studied by surface plasmon resonance. Biotinylated bungarotoxin, immobilized on a streptavidin-coated gold film, binds nicotinic acetylcholine receptor both in detergent-solubilized and in lipid vesicle-reconstituted form with high specificity. In the latter case, nonspecific binding to the sensor surface is significantly reduced by reconstituting the receptor into poly(ethylene glycol)-lipid-contg. sterically stabilized vesicles. By preincubation of a bulk nicotinic acetylcholine receptor sample with the competing ligands carbamoylcholine and decamethonium bromide, the subsequent specific binding of the receptor to the surface-immobilized bungarotoxin is reduced, depending on the concn. of competing ligand. This competition assay allows the detn. of the dissocn. consts. of the acetylcholine receptor-carbamoylcholine complex. A KD = 3.5 * 10-6 M for the detergent-solubilized receptor and a KD = 1.4 * 10-5 M for the lipid vesicle-reconstituted receptor are obtained. For decamethonium bromide, a KD = 4.5 * 10-5 M is detd. for the detergent-solubilized receptor. This approach is of general importance for investigating ligand-receptor interactions in case of small ligand mols. by mass-sensitive techniques. [on SciFinder (R)]

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