Pharmacological analysis of a dopamine D2Short:Gαofusion protein expressed in Sf9 cells
| Autor: | Lucien Gazi, Juan F. López-Giménez, Thierry Wurch, Petrus J. Pauwels, Philip G. Strange |
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| Rok vydání: | 2003 |
| Předmět: |
Spiperone
Apomorphine GTP' D2 dopamine receptor Stereochemistry Recombinant Fusion Proteins Biophysics GTPgammaS [3H]Spiperone binding Spodoptera Ligands Binding Competitive Biochemistry [35S]Guanosine-5′-O-(3-thio)triphosphate binding Cell Line chemistry.chemical_compound GTP-binding protein regulators GTP-Binding Proteins Structural Biology Genetics medicine Animals Humans Drug Interactions G protein-coupled receptor Baculovirus Binding site Molecular Biology Binding Sites Receptors Dopamine D2 Guanosine 5'-O-(3-Thiotriphosphate) Cell Biology Fusion protein Molecular biology chemistry Dopamine Agonists Dopamine Antagonists Baculoviridae medicine.drug |
| Zdroj: | FEBS Letters. 545:155-160 |
| ISSN: | 0014-5793 |
| Popis: | A dopamine D(2Short) receptor:G(alphao) fusion protein was expressed in Sf9 cells using the baculovirus expression system. [(3)H]Spiperone bound to D(2Short):G(alphao) with a pK(d) approximately 10. Dopamine stimulated the binding of [(35)S]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 [(35)S]GTPgammaS binding) at D(2Short):G(alphao)/Gbeta(1)gamma(2) although (-)-sulpiride and ziprasidone were neutral antagonists. Competition of [(3)H]spiperone binding by dopamine and 10,11-dihydroxy-N-n-propylnorapomorphine revealed two binding sites of different affinities, even in the presence of GTP (100 micro M). The D(2Short):G(alphao) fusion protein is therefore a good model for characterising D(2) receptors. |
| Databáze: | OpenAIRE |
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