Two isoforms of the EP3 receptor with different carboxyl-terminal domains. Identical ligand binding properties and different coupling properties with Gi proteins

• Published in: The Journal of biological chemistry 1993-02, Vol.268 (4), p.2712-2718
• Main Authors: SUGIMOTO, Y, NEGISHI, M, HAYASHI, Y, NAMBA, T, HONDA, A, WATABE, A, HIRATA, M, NARUMIYA, S, ICHIKAWA, A
• Format: Article
• Language: English
• Subjects: Adenylyl Cyclases - metabolism; Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Cell receptors; Cell structures and functions; Cloning, Molecular; Cytoplasm - metabolism; Dinoprostone - physiology; DNA - genetics; Fundamental and applied biological sciences. Psychology; GTP Phosphohydrolases - metabolism; GTP-Binding Proteins - metabolism; Guanosine 5'-O-(3-Thiotriphosphate) - metabolism; Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors; Ligands; Mice; Molecular and cellular biology; Molecular Sequence Data; Pertussis Toxin; Receptors, Prostaglandin - genetics; Receptors, Prostaglandin - metabolism; Receptors, Prostaglandin - ultrastructure; Receptors, Prostaglandin E; Restriction Mapping; RNA Splicing; Signal Transduction; Solubility; Virulence Factors, Bordetella - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)53832-1

Functional cDNA clones for two isoforms of the mouse prostaglandin E receptor EP3 subtype derived from alternative RNA splicing were obtained. The two isoforms are only different in the sequence of the putative cytoplasmic carboxyl-terminal tail and their hydrophobicity; one isoform, named EP3 alpha, has a hydrophilic tail, and the other, named EP3 beta, has a hydrophobic tail. When expressed, the two receptors displayed identical ligand binding properties but different responses to guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S). Without a change in the Bmax value, GTP gamma S increased Kd for prostaglandin E2 of EP3 beta and decreased that of EP3 alpha. These effects were abolished by the treatment of membranes with pertussis toxin and restored by the addition of Gi2. Although both isoforms exerted inhibition of forskolin-induced cAMP accumulation, three orders lower concentrations of agonists were required for EP3 alpha than EP3 beta for 50% inhibition of cAMP formation. A similar difference in agonist potency was observed also for agonist-induced stimulation of GTPase activity in membranes. Thus, the two receptors with different carboxyl-terminal tails show different coupling to the Gi protein, leading to the opposite responses to GTP in the ligand binding affinity and to different affinities of the agonist-occupied receptors to the G proteins.