A mu-opioid receptor-filter assay. Rapid estimation of binding affinity of ligands and reversibility of long-lasting ligand-receptor complexes.

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Ilien B, Galzi JL, Mejean A, Goeldner M, Hirth C

A mu-opioid receptor-filter assay. Rapid estimation of binding affinity of ligands and reversibility of long-lasting ligand-receptor complexes.

Biochem Pharmacol. 1988 Oct 15;37(20):3843-51.

PubMed ID

2847746 [ View in PubMed

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Abstract

A filter-associated binding technique, originally described by Leysen and Gommeren [J. Receptor Res. 4, 817 (1984); Drug Dev. Res. 8, 119 (1986)], was focused on the study of mu-opioid receptor sites. The interesting binding features of 3H-Sufentanil, a mu-selective radioligand, permitted such a filter procedure to be performed. Its application was 3-fold. A 5-min binding assay allowed us to verify that specific 3H-Sufentanil binding to filter-absorbed rat brain membranes was endowed with the known kinetic and equilibrium binding properties of membrane-bound mu-opioid receptor sites. Moreover, the filtration technique allowed us to get rapid information about the dissociation rates of unlabelled compounds from the receptor sites. In practice, membranes, preincubated with high concentrations of cold drugs, were absorbed to filters. Dissociation was achieved by repeatedly applying buffer samples on the filter, and monitored by the recovery in free specific 3H-Sufentanil binding sites. Finally, such a dissociation procedure, improved by a washing buffer at high ionic strength, was found to be much more efficient and attractive than the classical dilution-centrifugation procedure, especially for slowly-dissociating compounds. Special attention was paid to the discrimination between pseudo-irreversible binding of drugs and stable covalent labelling of mu-opioid receptor sites (either by affinity or photoaffinity probes), particularly when unlabelled ligands were the only tools available.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Sufentanil	Mu-type opioid receptor	Protein	Humans	Yes	Agonist	Details