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 Pho tinus pyral is luciferase into which a cAMP-binding protein moiety has been inserted. Results, expressed as EC50 or IC50 values for agonists and antagonists, respectively, showed a strong correlation with those obtained with [35S]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 Gαi.
Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17
BUCCIONI, Michela;MARUCCI, Gabriella;DAL BEN, DIEGO;GIACOBBE, Dania;LAMBERTUCCI, Catia;SOVERCHIA, Laura;THOMAS, AJIROGHENE;VOLPINI, Rosaria;CRISTALLI, Gloria
2011-01-01
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 Pho tinus pyral is luciferase into which a cAMP-binding protein moiety has been inserted. Results, expressed as EC50 or IC50 values for agonists and antagonists, respectively, showed a strong correlation with those obtained with [35S]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 Gαi.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
