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Pharmacological analysis of a dopamine D-2Short: G(alpha o) fusion protein expressed in Sf9 cells

Gazi, L., Wurch, T., Lopez-Gimenez, J. F., Pauwels, P. J. and Strange, P. G. (2003) Pharmacological analysis of a dopamine D-2Short: G(alpha o) fusion protein expressed in Sf9 cells. FEBS Letters, 545 (2-3). pp. 155-160. ISSN 0014-5793

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To link to this item DOI: 10.1016/s0014-5793(03)00520-9

Abstract/Summary

A dopamine D-2Short receptor:G(alphao) fusion protein was expressed in Sf9 cells using the baculovirus expression system. [H-3]Spiperone bound to D-2Short:G(alphao) with a pK(d) approximate to 10. Dopamine stimulated the binding of [S-35]guanosine-5'-O-(3-thio)triphosphate (GTPgammaS) to D-2Short:G(alphao) expressed with Gbeta(1)gamma(2) (E-max > 460%; pEC(50) 5.43 +/- 0.06). Most of the putative D-2 antagonists behaved as inverse agonists (suppressing basal [S-35]GTPgammaS binding) at D-2Short:G(alphao)/Gbeta(1)gamma(2) although (-)-suipiride and ziprasidone were neutral antagonists. Competition of [H-3]spiperone binding by dopamine and 10,11-dihydroxy-N-n-propylnorapo-morphine revealed two, binding sites of different affinities, even in the presence of GTP (100 muM). The D-2Short:G(alphao) fusion protein is therefore a good model for characterising D-2 receptors. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Biological Sciences
ID Code:10744
Uncontrolled Keywords:D-2 dopamine receptor, G protein-coupled receptor, fusion protein, baculovirus, [H-3]spiperone binding, [S-35]guanosine-5, '-O-('3-thio)triphosphate binding, SIGNAL-TRANSDUCTION, AGONIST REGULATION, MOLECULAR ANALYSIS, RECEPTOR, ACTIVATION, RECONSTITUTION, ANTAGONISTS, SPECIFICITY, SUBUNITS, GAMMA

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