Publication

G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli: Clues for a Constrained D3 Receptor Structure

Vanhauwe, J. F. M., Josson, K., Luyten, W. H. M. L., Driessen, A. J. & Leysen, J. E., 2000, In : Journal of Pharmacology and Experimental Therapeutics. 295, 1, p. 274-283 10 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Vanhauwe, J. F. M., Josson, K., Luyten, W. H. M. L., Driessen, A. J., & Leysen, J. E. (2000). G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli: Clues for a Constrained D3 Receptor Structure. Journal of Pharmacology and Experimental Therapeutics, 295(1), 274-283.

Author

Vanhauwe, Jurgen F.M. ; Josson, Katty ; Luyten, Walter H.M.L. ; Driessen, Arnold J. ; Leysen, Josée E. / G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli : Clues for a Constrained D3 Receptor Structure. In: Journal of Pharmacology and Experimental Therapeutics. 2000 ; Vol. 295, No. 1. pp. 274-283.

Harvard

Vanhauwe, JFM, Josson, K, Luyten, WHML, Driessen, AJ & Leysen, JE 2000, 'G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli: Clues for a Constrained D3 Receptor Structure', Journal of Pharmacology and Experimental Therapeutics, vol. 295, no. 1, pp. 274-283.

Standard

G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli : Clues for a Constrained D3 Receptor Structure. / Vanhauwe, Jurgen F.M.; Josson, Katty; Luyten, Walter H.M.L.; Driessen, Arnold J.; Leysen, Josée E.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 295, No. 1, 2000, p. 274-283.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Vanhauwe JFM, Josson K, Luyten WHML, Driessen AJ, Leysen JE. G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli: Clues for a Constrained D3 Receptor Structure. Journal of Pharmacology and Experimental Therapeutics. 2000;295(1):274-283.


BibTeX

@article{cd8cd3d231e747b79b0e302d3cfbbd47,
title = "G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli: Clues for a Constrained D3 Receptor Structure",
abstract = "Human dopamine D2 and D3 receptors were expressed in Chinese hamster ovary (CHO) and Escherichia coli cells to compare their ligand binding properties in the presence or absence of G-proteins and to analyze their ability to interact with Gi/o-proteins. Binding affinities of agonists (dopamine, 7-OH-DPAT, PD128907, lisuride) and antagonists/inverse agonists (haloperidol, risperidone, domperidone, spiperone, raclopride, nemonapride), measured using [125I]iodosulpride and [3H]7-OH-DPAT, were similar for hD3 receptors in E. coli and CHO cell membranes. Both agonists and antagonists showed 2- to 25-fold lower binding affinities at hD2 receptors in E. coli versus CHO cell membranes (measured with [3H]spiperone), but the rank order of potencies remained similar. Purported inverse agonists did not display higher affinities for G-protein-free receptors. In CHO membranes, GppNHp decreased high affinity agonist ([3H]7-OH-DPAT) binding at hD2 receptors but not at hD3 receptors. Also, [3H]7-OH-DPAT (nanomolar concentration range) binding was undetectable at hD2 but clearly measurable at hD3 receptors in E. coli membranes. Addition of a Gi/o-protein mix to E. coli membranes increased high affinity [3H]7-OH-DPAT binding in a concentration-dependent manner at hD2 and hD3 receptors; this effect was reversed by addition of GppNHp. The potency of the Gi/o-protein mix to reconstitute high affinity binding was similar for hD2 and hD3 receptors. Thus, agonist binding to D3 receptors is only slightly affected by G-protein uncoupling, pointing to a rigid receptor structure. Furthermore, we propose that the generally reported lower signaling capacity of D3 receptors (versus D2 receptors) is not due to its lower affinity for G-proteins but attributed to its lower capacity to activate these G-proteins.",
author = "Vanhauwe, {Jurgen F.M.} and Katty Josson and Luyten, {Walter H.M.L.} and Driessen, {Arnold J.} and Leysen, {Jos{\'e}e E.}",
note = "Relation: https://www.rug.nl/gbb/ date_submitted:2007 Rights: University of Groningen, Groningen Biomolecular Sciences and Biotechnology Institute",
year = "2000",
language = "English",
volume = "295",
pages = "274--283",
journal = "Journal of Pharmacology and Experimental Therapeutics",
issn = "0022-3565",
publisher = "WILLIAMS & WILKINS",
number = "1",

}

RIS

TY - JOUR

T1 - G-Protein Sensitivity of Ligand Binding to Human Dopamine D2 and D3 Receptors Expressed in Escherichia coli

T2 - Clues for a Constrained D3 Receptor Structure

AU - Vanhauwe, Jurgen F.M.

AU - Josson, Katty

AU - Luyten, Walter H.M.L.

AU - Driessen, Arnold J.

AU - Leysen, Josée E.

N1 - Relation: https://www.rug.nl/gbb/ date_submitted:2007 Rights: University of Groningen, Groningen Biomolecular Sciences and Biotechnology Institute

PY - 2000

Y1 - 2000

N2 - Human dopamine D2 and D3 receptors were expressed in Chinese hamster ovary (CHO) and Escherichia coli cells to compare their ligand binding properties in the presence or absence of G-proteins and to analyze their ability to interact with Gi/o-proteins. Binding affinities of agonists (dopamine, 7-OH-DPAT, PD128907, lisuride) and antagonists/inverse agonists (haloperidol, risperidone, domperidone, spiperone, raclopride, nemonapride), measured using [125I]iodosulpride and [3H]7-OH-DPAT, were similar for hD3 receptors in E. coli and CHO cell membranes. Both agonists and antagonists showed 2- to 25-fold lower binding affinities at hD2 receptors in E. coli versus CHO cell membranes (measured with [3H]spiperone), but the rank order of potencies remained similar. Purported inverse agonists did not display higher affinities for G-protein-free receptors. In CHO membranes, GppNHp decreased high affinity agonist ([3H]7-OH-DPAT) binding at hD2 receptors but not at hD3 receptors. Also, [3H]7-OH-DPAT (nanomolar concentration range) binding was undetectable at hD2 but clearly measurable at hD3 receptors in E. coli membranes. Addition of a Gi/o-protein mix to E. coli membranes increased high affinity [3H]7-OH-DPAT binding in a concentration-dependent manner at hD2 and hD3 receptors; this effect was reversed by addition of GppNHp. The potency of the Gi/o-protein mix to reconstitute high affinity binding was similar for hD2 and hD3 receptors. Thus, agonist binding to D3 receptors is only slightly affected by G-protein uncoupling, pointing to a rigid receptor structure. Furthermore, we propose that the generally reported lower signaling capacity of D3 receptors (versus D2 receptors) is not due to its lower affinity for G-proteins but attributed to its lower capacity to activate these G-proteins.

AB - Human dopamine D2 and D3 receptors were expressed in Chinese hamster ovary (CHO) and Escherichia coli cells to compare their ligand binding properties in the presence or absence of G-proteins and to analyze their ability to interact with Gi/o-proteins. Binding affinities of agonists (dopamine, 7-OH-DPAT, PD128907, lisuride) and antagonists/inverse agonists (haloperidol, risperidone, domperidone, spiperone, raclopride, nemonapride), measured using [125I]iodosulpride and [3H]7-OH-DPAT, were similar for hD3 receptors in E. coli and CHO cell membranes. Both agonists and antagonists showed 2- to 25-fold lower binding affinities at hD2 receptors in E. coli versus CHO cell membranes (measured with [3H]spiperone), but the rank order of potencies remained similar. Purported inverse agonists did not display higher affinities for G-protein-free receptors. In CHO membranes, GppNHp decreased high affinity agonist ([3H]7-OH-DPAT) binding at hD2 receptors but not at hD3 receptors. Also, [3H]7-OH-DPAT (nanomolar concentration range) binding was undetectable at hD2 but clearly measurable at hD3 receptors in E. coli membranes. Addition of a Gi/o-protein mix to E. coli membranes increased high affinity [3H]7-OH-DPAT binding in a concentration-dependent manner at hD2 and hD3 receptors; this effect was reversed by addition of GppNHp. The potency of the Gi/o-protein mix to reconstitute high affinity binding was similar for hD2 and hD3 receptors. Thus, agonist binding to D3 receptors is only slightly affected by G-protein uncoupling, pointing to a rigid receptor structure. Furthermore, we propose that the generally reported lower signaling capacity of D3 receptors (versus D2 receptors) is not due to its lower affinity for G-proteins but attributed to its lower capacity to activate these G-proteins.

M3 - Article

VL - 295

SP - 274

EP - 283

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

IS - 1

ER -

ID: 14463601