A chimeric D2 dopamine/m1 muscarinic receptor with D2 binding specificity mobilizes intracellular calcium in response to dopamine

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 m1 muscarinic receptor. When expressed in CHO cells, the chimeric D2/m1 receptor bound dopaminergic ligands...

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Bibliographic Details
Published inFEBS letters Vol. 279; no. 1; pp. 87 - 90
Main Authors ENGLAND, B. P, ACKERMAN, M. S, BARRETT, R. W
Format Journal Article
LanguageEnglish
Published Amsterdam Elsevier 11.02.1991
Subjects
Animals
Base Sequence
Biological and medical sciences
Calcium - metabolism
Cell Line
Cell receptors
Cell structures and functions
Chimera
Cricetinae
Cricetulus
Dopamine - pharmacology
Fundamental and applied biological sciences. Psychology
Fura-2
Humans
Miscellaneous
Molecular and cellular biology
Molecular Sequence Data
Radioligand Assay
Receptors, Dopamine - genetics
Receptors, Dopamine - metabolism
Receptors, Muscarinic - genetics
Receptors, Muscarinic - metabolism
Substrate Specificity
Transfection
Human
Ligand binding
Signal transduction
Heterologous system
Dopamine
D2 Dopamine receptor
Calcium
Scatchard plot
Structure function relationship
M1 receptor
Hybrid gene
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Summary: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 m1 muscarinic receptor. When expressed in CHO cells, the chimeric D2/m1 receptor bound dopaminergic ligands with affinities similar to the native D2(414) receptor. Intracellular calcium levels (measured with Fura-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/m1 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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(91)80257-4