Abstract
The human D4 dopamine receptor has been genetically engineered for expression in insect cells using the baculovirus system. A D4 cDNA gene fusion construct [(1991) Nature 350, 610-614] was synthetically modified to remove two introns from the coding region, and expressed in S. frugiperda (Sf9) cells as a fusion with a short sequence from the polyhedrin protein. Binding assays with [3H]spiperone indicated high levels of D4 receptor binding 90 h after infection and a pharmacological profile identical to that reported for D4 receptors expressed in COS-7 cells using the cDNA gene hybrid. We also show that the agonist binding affinity of D4 receptors expressed in Sf9 cells can be shifted by GTP-gamma-S, indicating coupling to G-proteins.
MeSH terms
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Animals
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Baculoviridae
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Base Sequence
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Cell Line
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Ergolines / metabolism
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GTP-Binding Proteins / metabolism
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Genetic Vectors
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Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
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Humans
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Introns
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Molecular Sequence Data
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Moths
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Occlusion Body Matrix Proteins
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Quinpirole
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Receptors, Dopamine / biosynthesis
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Receptors, Dopamine / genetics*
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Receptors, Dopamine / metabolism
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Receptors, Dopamine D2*
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Receptors, Dopamine D4
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Recombinant Fusion Proteins / biosynthesis*
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Recombinant Fusion Proteins / metabolism
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Spiperone / metabolism
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Viral Proteins / genetics
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Viral Structural Proteins
Substances
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DRD4 protein, human
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Ergolines
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Occlusion Body Matrix Proteins
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Receptors, Dopamine
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Receptors, Dopamine D2
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Recombinant Fusion Proteins
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Viral Proteins
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Viral Structural Proteins
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polyhedrin protein, Nucleopolyhedrovirus
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Receptors, Dopamine D4
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Quinpirole
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Guanosine 5'-O-(3-Thiotriphosphate)
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Spiperone
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GTP-Binding Proteins