PMID: 9159497Apr 25, 1997Paper

Agonist-induced [35S]GTPgammaS binding in the membranes of Spodoptera frugiperda insect cells expressing the human D3 dopamine receptor

Neuroscience Letters
J F PregenzerW B Im

Abstract

In the membranes of Spodoptera frugiperda (Sf-9) insect cells heterologously expressing the human D3 dopamine receptor, agonists selective for the receptor, but not antagonists, robustly enhanced [35S]GTPgammaS binding. Quinpirole, for instance, dose-dependently enhanced [35S]GTPgammaS binding with a half-maximal concentration of 2.3 +/- 0.2 nM. Its action was absent in the cells infected with wild type viruses, and competitively blocked by an antagonist, YM-09151-2. A number of known agonists enhanced [35S]GTPgammaS binding to variable degrees, probably reflecting their differential efficacy to activate target G-proteins via the receptor. This agonist-induced [35S]GTPgammaS binding was abolished by N-ethylmaleimide, a selective blocking agent for Gi/Go proteins, with no appreciable effect on ligand binding. We propose coupling of the cloned D3 receptor to endogenous G-proteins in Sf-9 cells, probably homologs of mammalian Gi/Go proteins. Despite the apparent coupling of the D3 receptor to G-proteins, GTPgammaS (10 microM) failed to decrease agonist binding ([3H]dopamine) to the D3 receptor, probably due to small affinity differences between low and high affinity states for agonists in the D3 receptor, as well as due to high re...Continue Reading

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Citations

Feb 5, 2003·Biochemical and Biophysical Research Communications·Teresa M KubiakJerry W Bowman
Sep 16, 2003·Neurochemistry International·Ain UustareAgo Rinken
Dec 17, 2014·European Neuropsychopharmacology : the Journal of the European College of Neuropsychopharmacology·S KasselH Stark
Nov 4, 2004·Journal of Receptor and Signal Transduction Research·Joy A Ahlgren-Beckendorf, Beth Levant

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