Expression and characterization of human D4 dopamine receptors in baculovirus-infected insect cells.
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.
Mills
A
A
Glaxo Institute for Molecular Biology SA, Plan-les-Ouates/Geneva, Switzerland.
Allet
B
B
Bernard
A
A
Chabert
C
C
Brandt
E
E
Cavegn
C
C
Chollet
A
A
Kawashima
E
E
D000818:Animals; D016367:Baculoviridae; D001483:Base Sequence; D002460:Cell Line; D004873:Ergolines / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D005822:Genetic Vectors; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009036:Moths; D000077183:Occlusion Body Matrix Proteins; D019257:Quinpirole; D011954:Receptors, Dopamine / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D017448:Receptors, Dopamine D2; D050638:Receptors, Dopamine D4; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000378:metabolism; D013134:Spiperone / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins
C493599:DRD4 protein, human; D004873:Ergolines; D000077183:Occlusion Body Matrix Proteins; D011954:Receptors, Dopamine; D017448:Receptors, Dopamine D2; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D050638:Receptors, Dopamine D4; D019257:Quinpirole; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D013134:Spiperone; D019204:GTP-Binding Proteins