Structural insights into the G protein selectivity revealed by the human EP3-Gi signaling complex

Ryoji Suno; Yukihiko Sugita; Kazushi Morimoto; Hiroko Takazaki; Hirokazu Tsujimoto; Mika Hirose; Chiyo Suno-Ikeda; Norimichi Nomura; Tomoya Hino; Asuka Inoue; Kenji Iwasaki; Takayuki Kato; So Iwata; Takuya Kobayashi

doi:10.1016/j.celrep.2022.111323

Structural insights into the G protein selectivity revealed by the human EP3-G_i signaling complex

Cell Rep. 2022 Sep 13;40(11):111323. doi: 10.1016/j.celrep.2022.111323.

Authors

Ryoji Suno¹, Yukihiko Sugita², Kazushi Morimoto³, Hiroko Takazaki⁴, Hirokazu Tsujimoto⁵, Mika Hirose⁴, Chiyo Suno-Ikeda⁶, Norimichi Nomura⁵, Tomoya Hino⁷, Asuka Inoue⁸, Kenji Iwasaki⁹, Takayuki Kato⁴, So Iwata⁵, Takuya Kobayashi¹⁰

Affiliations

¹ Department of Medical Chemistry, Kansai Medical University, Hirakata 573-1010, Japan. Electronic address: sunory@hirakata.kmu.ac.jp.
² Institute for Protein Research, Osaka University, Suita 565-0871, Japan; Hakubi Center for Advanced Research, Kyoto University, Kyoto 606-8501, Japan; Institute for Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan.
³ Physical Chemistry for Life Science Laboratory, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
⁴ Institute for Protein Research, Osaka University, Suita 565-0871, Japan.
⁵ Department of Cell Biology and Medical Chemistry, Graduate School of Medicine, Kyoto University, Konoe-cho, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan.
⁶ Department of Medical Chemistry, Kansai Medical University, Hirakata 573-1010, Japan.
⁷ Department of Chemistry and Biotechnology, Graduate School of Engineering, Tottori University, Tottori, Japan; Center for Research on Green Sustainable Chemistry, Tottori University, Tottori, Japan.
⁸ Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3, Aoba, Aramaki, Aoba-Ku, Sendai, Miyagi 980-8578, Japan.
⁹ Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, Tsukuba, Japan.
¹⁰ Department of Medical Chemistry, Kansai Medical University, Hirakata 573-1010, Japan; Japan Agency for Medical Research and Development (AMED), Core Research for Evolutional Science and Technology (CREST), 1-7-1 Otemachi, Chiyoda-ku, Tokyo 100-0004, Japan. Electronic address: kobayatk@hirakata.kmu.ac.jp.

PMID: 36103815
DOI: 10.1016/j.celrep.2022.111323

Abstract

Prostaglandin receptors have been implicated in a wide range of functions, including inflammation, immune response, reproduction, and cancer. Our group has previously determined the crystal structure of the active-like EP3 bound to its endogenous agonist, prostaglandin E₂. Here, we present the single-particle cryoelectron microscopy (cryo-EM) structure of the human EP3-G_i signaling complex at a resolution of 3.4 Å. The structure reveals the binding mode of G_i to EP3 and the structural changes induced in EP3 by G_i binding. In addition, we compare the structure of the EP3-G_i complex with other subtypes of prostaglandin receptors (EP2 and EP4) bound to G_s that have been previously reported and examine the differences in amino acid composition at the receptor-G protein interface. Mutational analysis reveals that the selectivity of the G protein depends on specific amino acid residues in the second intracellular loop and TM5.

Keywords: CP: Molecular biology; G protein-coupled receptor; G protein-coupling selectivity; cryoelectron microscopy single-particle analysis; prostaglandin receptor; signal transduction.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acids
Cryoelectron Microscopy
Dinoprostone* / pharmacology
Humans
Receptors, Prostaglandin E* / agonists
Receptors, Prostaglandin E* / metabolism
Receptors, Prostaglandin E, EP3 Subtype / metabolism

Substances

Amino Acids
Receptors, Prostaglandin E
Receptors, Prostaglandin E, EP3 Subtype
Dinoprostone