title: Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17.
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).
authors.affiliation: Medicinal Chemistry Unit, School of Pharmacy, University of Camerino, 62032, Camerino, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Buccioni
authors.forename: Gabriella
authors.initials: G
authors.lastname: Marucci
authors.forename: Diego
authors.initials: D
authors.lastname: Dal Ben
authors.forename: Dania
authors.initials: D
authors.lastname: Giacobbe
authors.forename: Catia
authors.initials: C
authors.lastname: Lambertucci
authors.forename: Laura
authors.initials: L
authors.lastname: Soverchia
authors.forename: Ajiroghene
authors.initials: A
authors.lastname: Thomas
authors.forename: Rosaria
authors.initials: R
authors.lastname: Volpini
authors.forename: Gloria
authors.initials: G
authors.lastname: Cristalli