A chimeric D2 dopamine/ml muscarinic receptor with D2 binding specificity mobilizes intracellular calcium in response to dopamine

FEBS Letters

Barrett, Ronald W.¹  England, Bruce P.¹  Ackerman, Martha S.¹ 

[1]Affymax Research Institute, 3180 Porter Drive, Palo Alto, CA 94304, USA

关键词: Dopamine receptor;    Muscarinic receptor;    Chimeric receptor;    Effector coupling;    Calcium mobilization;    QNB;    quinuclidinyl benzilate;    [Ca+2]1;    intracellular free calcium concentration;    PCR;    polymerase chain reaction;    CHO;    Chinese hamster ovary;   

DOI  :  10.1016/0014-5793(91)80257-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using PCR methodology, a chimeric receptor cDNA was constructed in which the entire third cytoplasmic loop of the human D₂ dopamine receptor was replaced by the analogous portion of the human ml muscarinic receptor. When expressed in CHO cells, the chimeric D₂/ml receptor bound dopaminergic ligands with affinities similar to the native D₂₍₄₁₄₎ receptor. Intracellular calcium levels (measured with Furu-2) were not altered when CHO cells expressing the D₂₍₄₁₄₎ receptor were exposed to dopamine. In contrast, dopamine elevated intracellular calcium levels in cells expressing the D₂/ml chimeric receptor in a dose-dependent manner which was blocked by the D₂ antagonist, fluphenazine. The ability to construct G-protein-linked receptor chimeras which mobilize calcium with nearly unaltered pharmacologic specificity raises the possibility of a generic strategy for creating non-radioisotopic reporter systems for use in drug discovery.

【 授权许可】

Unknown