Two isoforms of the prostaglandin E receptor EP3 subtype different in agonist-independent constitutive activity.
We previously identified two isoforms of the mouse prostaglandin E receptor EP3 subtype, EP3 alpha and EP3 beta, with different carboxyl-terminal tails, produced through alternative splicing and showing different efficiency in inhibition of adenylate cyclase (Sugimoto, Y., Negishi, M., Hayashi, Y., Namba, T., Honda, A., Watabe, A., Hirata, M., Narumiya, S., and Ichikawa, A. (1993) J. Biol. Chem. 268, 2712-2718). To assess the role of the carboxyl-terminal tails in the G protein coupling properties of the EP3 receptor, we examined the Gi activities of EP3 alpha, EP3 beta, and the mutant receptor, in which the carboxyl-terminal tail was truncated at the splicing site. The EP3 alpha receptor showed marked agonist-independent constitutive inhibition of adenylate cyclase, while EP3 beta receptor had no agonist-independent inhibition. On the other hand, the truncated receptor showed only agonist-independent constitutive inhibition. The constitutive activity of these receptors on the stimulation of GTPase activity of Gi was also observed. Thus, alternative splicing produced two isoforms with different carboxyl-terminal tails and with different constitutive activity, and the truncation of the carboxyl-terminal tail caused full constitutive activity.
Hasegawa
H
H
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Negishi
M
M
Ichikawa
A
A
D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D000527:Alprostadil / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D003593:Cytoplasm / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D018078:Receptors, Prostaglandin E / Q000502:physiology*; D011994:Recombinant Proteins; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D018078:Receptors, Prostaglandin E; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; C032405:11-deoxy-16-phenoxy-17,18,19,20-tetranorprostaglandin E1; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D000527:Alprostadil