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Development of tolerance in D3 dopamine receptor signaling is accompanied by distinct changes in receptor conformation
Journal article   Peer reviewed

Development of tolerance in D3 dopamine receptor signaling is accompanied by distinct changes in receptor conformation

Ligia Westrich, Sara Gil-Mast, Sandhya Kortagere and Eldo V Kuzhikandathil
Biochemical pharmacology, v 79(6), pp 897-907
2010
PMID: 19879251

Abstract

Signaling GPCR Desensitization Biotinylation Drd3 Modeling
The D3 dopamine receptor tolerance property develops after the removal of the agonist and is accompanied by distinct changes in receptor conformation which uncouples the receptor from its signaling cascade. The D3 but not D2 dopamine receptors exhibit a tolerance property in which agonist-induced D3 receptor response progressively decreases upon repeated agonist stimulation. We have previously shown that the D3 receptor tolerance property is not mediated by receptor internalization, persistent agonist binding or a decrease in receptor binding affinity. In this paper, we test the hypothesis that alterations in D3 receptor conformation underlie the tolerance property. Structural models of wild type and mutant human D3 receptors were generated using the beta adrenergic receptor crystal structure as a template. These models suggested that the agonist-bound D3 receptor undergoes conformational changes that could underlie its tolerance property. To experimentally assess changes in receptor conformation, we measured the accessibility of native cysteine residues present in the extracellular and transmembrane regions of the human D3 receptor to two different thiol-modifying biotinylating reagents. The accessibilities of the native cysteine residues present in the D3 receptor were assessed under control conditions, in the presence of agonist and under conditions that induced receptor tolerance. By comparing the accessibility of D3 receptor cysteine residues to hydrophobic and hydrophilic thiol-modifying biotinylating reagents, we show that the alteration of D3 receptor conformation during tolerance involves the net movement of cysteine residues into a hydrophobic environment. Our results show that the conformation state of the D3 receptor during tolerance is distinct from the conformation under basal and agonist-bound conditions. The results suggest that the D3 receptor tolerance property is mediated by conformational changes that may uncouple the receptor from G-protein signaling.

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Collaboration types
Domestic collaboration
Web of Science research areas
Pharmacology & Pharmacy
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