Published in
Springer Verlag, Purinergic Signalling, 4(7), p. 463-468
DOI: 10.1007/s11302-011-9245-8
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- ORCID
- ORCID
- Springer Verlag
- [europepmc.org] | PDF
- [www.researchgate.net] | PDF
- [www.ncbi.nlm.nih.gov] | PDF
- [www.ncbi.nlm.nih.gov] | PDF
Tools
Innovative functional cAMP assay for studying G protein-coupled receptors: Application to the pharmacological characterization of GPR17
,
Dania Giacobbe,
Catia Lambertucci,
Laura Soverchia,
Ajiroghene Thomas ,
Rosaria Volpini,
Gloria Cristalli
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Abstract
In this work, an innovative and non-radioactive functional cAMP assay was validated at the GPR17 receptor. This assay provides a simple and powerful new system to monitor G protein-coupled receptor activity through change in the intracellular cAMP concentration by using a mutant form of Photinus pyralis luciferase into which a cAMP-binding protein moiety has been inserted. Results, expressed as EC(50) or IC(50) values for agonists and antagonists, respectively, showed a strong correlation with those obtained with [(35)S]GTPγS binding assay, thus confirming the validity of this approach in the study of new ligands for GPR17. Moreover, this method allowed confirming that GPR17 is coupled with a G(αi).