Cryo-EM structure of an activated GPR4–Gs signaling complex
Abstract G protein-coupled receptor 4 (GPR4) belongs to the subfamily of proton-sensing GPCRs (psGPCRs), which detect pH changes in extracellular environment and regulate diverse physiological responses. GPR4 was found to be overactivated in acidic tumor microenvironment as well as inflammation site...
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| Format: | Article |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-55901-2 |
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| author | Yitong Ma Yijie Wang Mengyuan Tang Yuan Weng Ying Chen Yueming Xu Shuxiao An Yiran Wu Suwen Zhao Huanhuan Xu Dali Li Mingyao Liu Weiqiang Lu Heng Ru Gaojie Song |
| author_facet | Yitong Ma Yijie Wang Mengyuan Tang Yuan Weng Ying Chen Yueming Xu Shuxiao An Yiran Wu Suwen Zhao Huanhuan Xu Dali Li Mingyao Liu Weiqiang Lu Heng Ru Gaojie Song |
| author_sort | Yitong Ma |
| collection | DOAJ |
| description | Abstract G protein-coupled receptor 4 (GPR4) belongs to the subfamily of proton-sensing GPCRs (psGPCRs), which detect pH changes in extracellular environment and regulate diverse physiological responses. GPR4 was found to be overactivated in acidic tumor microenvironment as well as inflammation sites, with a triad of acidic residues within the transmembrane domain identified as crucial for proton sensing. However, the 3D structure remains unknown, and the roles of other conserved residues within psGPCRs are not well understood. Here we report cryo-electron microscopy (cryo-EM) structures of active zebrafish GPR4 at both pH 6.5 and 8.5, each highlighting a distribution of histidine and acidic residues at the extracellular region. Cell-based assays show that these ionizable residues moderately influence the proton-sensing capacity of zebrafish GPR4, compared to the more significant effects of the triad residues. Furthermore, we reveal a cluster of aromatic residues within the orthosteric pocket that may propagate the signaling to the intercellular region via repacking the aromatic patch at the central region. This study provides a framework for future signaling and functional investigation of psGPCRs. |
| format | Article |
| id | doaj-art-22124521b3754038859e4c8dccc64e5b |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-22124521b3754038859e4c8dccc64e5b2025-01-12T12:29:41ZengNature PortfolioNature Communications2041-17232025-01-0116111010.1038/s41467-025-55901-2Cryo-EM structure of an activated GPR4–Gs signaling complexYitong Ma0Yijie Wang1Mengyuan Tang2Yuan Weng3Ying Chen4Yueming Xu5Shuxiao An6Yiran Wu7Suwen Zhao8Huanhuan Xu9Dali Li10Mingyao Liu11Weiqiang Lu12Heng Ru13Gaojie Song14Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityLife Sciences Institute, Second Affiliated Hospital of Zhejiang University School of Medicine, Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Zhejiang UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityiHuman Institute, ShanghaiTech UniversityiHuman Institute, ShanghaiTech UniversityCollege of Science, Yunnan Agricultural UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityLife Sciences Institute, Second Affiliated Hospital of Zhejiang University School of Medicine, Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Zhejiang UniversityShanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal UniversityAbstract G protein-coupled receptor 4 (GPR4) belongs to the subfamily of proton-sensing GPCRs (psGPCRs), which detect pH changes in extracellular environment and regulate diverse physiological responses. GPR4 was found to be overactivated in acidic tumor microenvironment as well as inflammation sites, with a triad of acidic residues within the transmembrane domain identified as crucial for proton sensing. However, the 3D structure remains unknown, and the roles of other conserved residues within psGPCRs are not well understood. Here we report cryo-electron microscopy (cryo-EM) structures of active zebrafish GPR4 at both pH 6.5 and 8.5, each highlighting a distribution of histidine and acidic residues at the extracellular region. Cell-based assays show that these ionizable residues moderately influence the proton-sensing capacity of zebrafish GPR4, compared to the more significant effects of the triad residues. Furthermore, we reveal a cluster of aromatic residues within the orthosteric pocket that may propagate the signaling to the intercellular region via repacking the aromatic patch at the central region. This study provides a framework for future signaling and functional investigation of psGPCRs.https://doi.org/10.1038/s41467-025-55901-2 |
| spellingShingle | Yitong Ma Yijie Wang Mengyuan Tang Yuan Weng Ying Chen Yueming Xu Shuxiao An Yiran Wu Suwen Zhao Huanhuan Xu Dali Li Mingyao Liu Weiqiang Lu Heng Ru Gaojie Song Cryo-EM structure of an activated GPR4–Gs signaling complex Nature Communications |
| title | Cryo-EM structure of an activated GPR4–Gs signaling complex |
| title_full | Cryo-EM structure of an activated GPR4–Gs signaling complex |
| title_fullStr | Cryo-EM structure of an activated GPR4–Gs signaling complex |
| title_full_unstemmed | Cryo-EM structure of an activated GPR4–Gs signaling complex |
| title_short | Cryo-EM structure of an activated GPR4–Gs signaling complex |
| title_sort | cryo em structure of an activated gpr4 gs signaling complex |
| url | https://doi.org/10.1038/s41467-025-55901-2 |
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