Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif

Abstract Endothelin type B receptor (ETBR) plays a crucial role in regulating blood pressure and humoral homeostasis, making it an important therapeutic target for related diseases. ETBR activation by the endogenous peptide hormones endothelin (ET)−1–3 stimulates several signaling pathways, includin...

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Main Authors: Kazutoshi Tani, Saori Maki-Yonekura, Ryo Kanno, Tatsuki Negami, Tasuku Hamaguchi, Malgorzata Hall, Akira Mizoguchi, Bruno M. Humbel, Tohru Terada, Koji Yonekura, Tomoko Doi
Format: Article
Language:English
Published: Nature Portfolio 2024-10-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-024-06905-z
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author Kazutoshi Tani
Saori Maki-Yonekura
Ryo Kanno
Tatsuki Negami
Tasuku Hamaguchi
Malgorzata Hall
Akira Mizoguchi
Bruno M. Humbel
Tohru Terada
Koji Yonekura
Tomoko Doi
author_facet Kazutoshi Tani
Saori Maki-Yonekura
Ryo Kanno
Tatsuki Negami
Tasuku Hamaguchi
Malgorzata Hall
Akira Mizoguchi
Bruno M. Humbel
Tohru Terada
Koji Yonekura
Tomoko Doi
author_sort Kazutoshi Tani
collection DOAJ
description Abstract Endothelin type B receptor (ETBR) plays a crucial role in regulating blood pressure and humoral homeostasis, making it an important therapeutic target for related diseases. ETBR activation by the endogenous peptide hormones endothelin (ET)−1–3 stimulates several signaling pathways, including Gs, Gi/o, Gq/11, G12/13, and β-arrestin. Although the conserved NPxxY motif in transmembrane helix 7 (TM7) is important during GPCR activation, ETBR possesses the lesser known NPxxL motif. In this study, we present the cryo-EM structure of the ETBR–Gi complex, complemented by MD simulations and functional studies. These investigations reveal an unusual movement of TM7 to the intracellular side during ETBR activation and the essential roles of the diverse NPxxL motif in stabilizing the active conformation of ETBR and organizing the assembly of the binding pocket for the α5 helix of Gi protein. These findings enhance our understanding of the interactions between GPCRs and G proteins, thereby advancing the development of therapeutic strategies.
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spelling doaj-art-e309a935434d4d2dbb4055a9ce1118fc2025-08-20T02:17:46ZengNature PortfolioCommunications Biology2399-36422024-10-017111310.1038/s42003-024-06905-zStructure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motifKazutoshi Tani0Saori Maki-Yonekura1Ryo Kanno2Tatsuki Negami3Tasuku Hamaguchi4Malgorzata Hall5Akira Mizoguchi6Bruno M. Humbel7Tohru Terada8Koji Yonekura9Tomoko Doi10Center for Computational Sciences, University of Tsukuba, 1-1-1 Tennodai, TsukubaRIKEN SPring-8 Center, 1-1-1, Kouto, SayoScientific Imaging Section, Research Support Division, Okinawa Institute of Science and Technology Graduate University (OIST), 1919-1, Tancha, Onna-son, Kunigami-gunDepartment of Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-kuRIKEN SPring-8 Center, 1-1-1, Kouto, SayoScientific Imaging Section, Research Support Division, Okinawa Institute of Science and Technology Graduate University (OIST), 1919-1, Tancha, Onna-son, Kunigami-gunGraduate School of Medicine, Mie University, 2–174 Edobashi TsuProvost Office, Okinawa Institute of Science and Technology Graduate University (OIST), 1919-1, Tancha, Onna-son, Kunigami-gunDepartment of Biotechnology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-kuRIKEN SPring-8 Center, 1-1-1, Kouto, SayoGraduate School of Science, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-kuAbstract Endothelin type B receptor (ETBR) plays a crucial role in regulating blood pressure and humoral homeostasis, making it an important therapeutic target for related diseases. ETBR activation by the endogenous peptide hormones endothelin (ET)−1–3 stimulates several signaling pathways, including Gs, Gi/o, Gq/11, G12/13, and β-arrestin. Although the conserved NPxxY motif in transmembrane helix 7 (TM7) is important during GPCR activation, ETBR possesses the lesser known NPxxL motif. In this study, we present the cryo-EM structure of the ETBR–Gi complex, complemented by MD simulations and functional studies. These investigations reveal an unusual movement of TM7 to the intracellular side during ETBR activation and the essential roles of the diverse NPxxL motif in stabilizing the active conformation of ETBR and organizing the assembly of the binding pocket for the α5 helix of Gi protein. These findings enhance our understanding of the interactions between GPCRs and G proteins, thereby advancing the development of therapeutic strategies.https://doi.org/10.1038/s42003-024-06905-z
spellingShingle Kazutoshi Tani
Saori Maki-Yonekura
Ryo Kanno
Tatsuki Negami
Tasuku Hamaguchi
Malgorzata Hall
Akira Mizoguchi
Bruno M. Humbel
Tohru Terada
Koji Yonekura
Tomoko Doi
Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif
Communications Biology
title Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif
title_full Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif
title_fullStr Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif
title_full_unstemmed Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif
title_short Structure of endothelin ETB receptor–Gi complex in a conformation stabilized by unique NPxxL motif
title_sort structure of endothelin etb receptor gi complex in a conformation stabilized by unique npxxl motif
url https://doi.org/10.1038/s42003-024-06905-z
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