title: Functional selectivity of natural and synthetic prostaglandin EP4 receptor ligands.
abstract: Classically, the prostaglandin E(2) (PGE(2)) receptor EP(4) has been classified as coupling to the Galpha(s) subunit, leading to intracellular cAMP increases. However, EP(4) signaling has been revealed to be more complex and also involves coupling to pertussis toxin-sensitive Galpha(i) proteins and beta-arrestin-mediated effects. There are now many examples of selective activation of independent pathways by G protein-coupled receptor (GPCR) ligands, a concept referred to as functional selectivity. Because most EP(4) ligands had thus far only been functionally characterized by their ability to stimulate cAMP production, we systematically determined the potencies and efficacies of a panel of EP(4) ligands for activation of Galpha(s), Galpha(i), and beta-arrestin relative to the endogenous ligand PGE(2). For this purpose, we adapted three bioluminescence resonance energy transfer (BRET) assays to evaluate the respective pathways in living cells. Our results suggest considerable functional selectivity among the tested, structurally related agonists. PGE(2) was the most selective in activating Galpha(s), whereas PGF(2alpha) and PGE(1) alcohol were the most biased for activating Galpha(i1) and beta-arrestin, respectively. We observed reversal in order of potencies between beta-arrestin 2 and Galpha(i1) functional assays comparing PGE(1) alcohol and either PGF(2alpha), PGD(2), or 7-[(1R,2R)-2-[(E,3R)-3-hydroxy-4-(phenoxy)but-1-enyl]-5-oxocyclopentyl]heptanoic acid (M&B28767). Most ligands were full agonists for the three pathways tested. Our results have implications for the use of PGE(2) analogs in experimental and possibly clinical settings, because their activity spectra on EP(4) differ from that of the native agonist. The BRET-based methodology used for this first systematic assessment of a set of EP(4) agonists should be applicable for the study of other GPCRs.
authors: Department of Biochemistry, Université de Montréal, Québec, Canada.
authors: Martin
authors: M
authors: Leduc
authors: Billy
authors: B
authors: Breton
authors: Céline
authors: C
authors: Galés
authors: Christian
authors: C
authors: Le Gouill
authors: Michel
authors: M
authors: Bouvier
authors: Sylvain
authors: S
authors: Chemtob
authors: Nikolaus
authors: N
authors: Heveker
mesh_terms: D000818:Animals; D019390:Arrestins / Q000378:metabolism / Q000502:physiology; D002460:Cell Line; D015232:Dinoprostone / Q000378:metabolism* / Q000502:physiology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism / Q000502:physiology; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007313:Insecta; D008024:Ligands; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D018078:Receptors, Prostaglandin E / Q000378:metabolism* / Q000502:physiology; D058309:Receptors, Prostaglandin E, EP4 Subtype; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
chemical_list: C000605254:ARRB2 protein, human; D019390:Arrestins; D008024:Ligands; C545650:PTGER4 protein, human; D018078:Receptors, Prostaglandin E; D058309:Receptors, Prostaglandin E, EP4 Subtype; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D019205:GTP-Binding Protein alpha Subunits, Gs; D015232:Dinoprostone