3 research outputs found
Molecular cloning and expression of a prostaglandin E2 receptor of the EP3β subtype from rat hepatocytes
AbstractRat hepatocytes have previously been reported to possess prostaglandin E2 receptors of the EP3-type (EP3-receptors) that inhibit glucagon-stimulated glycogenolysis by decreasing cAMP. Here, the isolation of a functional EP3β receptor cDNA clone from a rat hepatocyte cDNA library is reported. This clone can be translated into a 362-amino-acid protein, that displays over 95% homology to the EP3β receptor from mouse mastocytoma. The amino- and carboxy-terminal region of the protein are least conserved. Transiently transfected HEK 293 cells expressed a single binding site for PGE2 with an apparent Kd of 15 nM. PGE2 > PGF2α > PGD2 competed for [3H]PGE2 binding sites as did the EP3 receptor agonists M&B 28767 = sulprostone > misoprostol but not the EP1 receptor antagonist SC 19220. In stably transfected CHO cells M&B 28767 > sulprostone = PGE2 > misoprostol > PGF2α inhibited the forskolin-elicited cAMP formation. Thus, the characteristics of the EP3β receptor of rat hepatocytes closely resemble those of the EP3β receptor of mouse mastocytoma
The C-terminal domain of the human EP4 receptor confers agonist-induced receptor desensitization in a receptor hybrid with the rat EP3β receptor
AbstractProstaglandin E2 receptors (EPR), which belong to the family of heterotrimeric G protein-coupled ectoreceptors with seven transmembrane domains, can be classified into four subtypes according to their ligand binding and G protein coupling specificity. Of these, EP3βR is coupled to Gi, whereas EP4R is coupled to Gs. EP4R, in contrast to EP3βR, shows agonist-induced desensitization. The C-terminal domain and the third intracellular loop of these receptors have been implicated in G protein coupling specificity and desensitization. Here, receptor hybrids consisting of the main portion of rat EP3βR and either the C-terminal domain or the third intracellular loop of human EP4R were used to study the contribution of the respective receptor domains to G protein coupling and desensitization. Neither the EP4R C-terminal domain nor the EP4R third intracellular loop alone was sufficient to change the coupling specificity of the rEP3hEP4 receptor hybrids from Gi to Gs or to confer additional Gs coupling. However, the EP4R C-terminal domain but not the third intracellular loop was necessary and sufficient to mediate rapid agonist-induced, second messenger-independent desensitization in the Gi-coupled hybrid receptors