Expression and characterization of human D4 dopamine receptors in baculovirus‐infected insect cells

FEBS Letters

Allet, Bernard¹  Kawashima, Eric¹  Bernard, Alain¹  Mills, Ann¹  Cavegn, Catherine¹  Brandt, Emmanuelle¹  Chabert, Christian¹  Chollet, Angelika¹ 

[1] Glaxo Institute for Molecular Biology SA, 14 chemin des Aulx, 1228 Plan-les-Ouates/Geneva, Switzerland

关键词: D4 dopamine receptor;    Schizophrenia;    Baculovirus expression;    G-protein;    GTP-γ-S;    guanosine-5'-γ-thiotriphosphate;    MOI;    multiplicity of infection;    Sf9 cells;    Spodoptera frugiperda cells;   

DOI  :  10.1016/0014-5793(93)80077-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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-γ-S, indicating coupling to G-proteins.

【 授权许可】

Unknown