Studies on the Structure of the Ligand-Binding Site of the Brain D1-Dopamine Receptor

Hollis, Clare M. and Strange, Philip G. (1992) Studies on the Structure of the Ligand-Binding Site of the Brain D1-Dopamine Receptor. Biochemical Pharmacology, 44 (2). pp. 325-334. ISSN 0006-2952. (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1016/0006-2952(92)90016-C

Abstract

A series of group-specific modifying reagents were tested for their effects on [H-3]SCH23390 binding to brain D1 dopamine receptors in order to identify amino acid residues at the ligand binding site of the D1 dopamine receptor that are critical for ligand binding. Thc dependence of ligand binding on the pH of the incubation medium was also examined. The histidine-selective reagent, diethylpyrocarbonate did affect ligand binding but this is probably not due to an effect at the ligand binding site. Experiments with N-acetylimidazole and ethylacetimidate indicated that modification of tyrosine and amino residues did not exert major influences at the ligand binding site. The use of the thiol dithiothreitol indicated that breakage of a disulphide bond altered ligand binding, probably by affecting the receptor conformation, and the use of the sulphydryl reagent 5,5'-dithio-bis-nitrobenzoic acid showed that modification of a sulphydryl group on the receptor inhibited ligand binding. The carboxyl reagent N,N'-dicyclohexyl carbodiimide (DCCD) potently inhibited ligand binding and the effect could be prevented by occupancy of the receptor site by an agonist or antagonist so that there is an important carboxyl group at the receptor binding site. The total number of D1 receptors was reduced after the modification by DCCD and 70% of the residual receptors showed a reduced affinity for binding [H-3]SCH23390, the remainder having the same affinity as untreated receptors. [H-3]SCH23390 binding is also reduced by a decrease of pH and this effect seems to depend on the protonation of a group of pK(a) 6.9. Saturation analysis of [H-3]SCH23390 binding performed at pH 7.5 shows a single class of high affinity sites whereas at pH 6.0, two classes of sites with higher and lower affinities are seen. These studies suggested a model whereby [H-3]SCH23390 binding is to two receptor isoforms with different pH dependencies for [H-3]SCH23390 binding.

Item Type: Article
Subjects: R Medicine > RM Therapeutics. Pharmacology
Divisions: Faculties > Science Technology and Medical Studies > Medway School of Pharmacy
Depositing User: P. Ogbuji
Date Deposited: 07 Sep 2009 09:46
Last Modified: 17 Apr 2014 08:24
Resource URI: http://kar.kent.ac.uk/id/eprint/22617 (The current URI for this page, for reference purposes)
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