Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex
Abstract Unraveling the signaling roles of intermediate complexes is pivotal for G protein-coupled receptor (GPCR) drug development. Despite hundreds of GPCR-Gαβγ structures, these snapshots primarily capture the fully activated complex. Consequently, the functions of intermediate GPCR-G protein com...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56434-4 |
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| author | Maxine Bi Xudong Wang Jinan Wang Jun Xu Wenkai Sun Victor Ayo Adediwura Yinglong Miao Yifan Cheng Libin Ye |
| author_facet | Maxine Bi Xudong Wang Jinan Wang Jun Xu Wenkai Sun Victor Ayo Adediwura Yinglong Miao Yifan Cheng Libin Ye |
| author_sort | Maxine Bi |
| collection | DOAJ |
| description | Abstract Unraveling the signaling roles of intermediate complexes is pivotal for G protein-coupled receptor (GPCR) drug development. Despite hundreds of GPCR-Gαβγ structures, these snapshots primarily capture the fully activated complex. Consequently, the functions of intermediate GPCR-G protein complexes remain elusive. Guided by a conformational landscape visualized via 19F quantitative NMR and molecular dynamics (MD) simulations, we determined the structure of an intermediate GPCR-mini-Gαsβγ complex at 2.6 Å using cryo-EM, by blocking its transition to the fully activated complex. Furthermore, we present direct evidence that the complex at this intermediate state initiates a rate-limited nucleotide exchange before transitioning to the fully activated complex. In this state, BODIPY-GDP/GTP based nucleotide exchange assays further indicated the α-helical domain of the Gα is partially open, allowing it to grasp a nucleotide at a non-canonical binding site, distinct from the canonical nucleotide-binding site. These advances bridge a significant gap in our understanding of the complexity of GPCR signaling. |
| format | Article |
| id | doaj-art-493d69db7b6e44b481428359157c03b3 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-493d69db7b6e44b481428359157c03b32025-02-02T12:33:06ZengNature PortfolioNature Communications2041-17232025-01-0116111410.1038/s41467-025-56434-4Structure and function of a near fully-activated intermediate GPCR-Gαβγ complexMaxine Bi0Xudong Wang1Jinan Wang2Jun Xu3Wenkai Sun4Victor Ayo Adediwura5Yinglong Miao6Yifan Cheng7Libin Ye8Department of Biochemistry and Biophysics, University of CaliforniaDepartment of Molecular Biosciences, University of South Florida, 4202 E Fowler AvePharmacology & Computational Medicine Program, University of North Carolina at Chapel Hill, 116 Manning DriveDepartment of Molecular and Cellular Physiology, Stanford University School of MedicineDepartment of Molecular Biosciences, University of South Florida, 4202 E Fowler AvePharmacology & Computational Medicine Program, University of North Carolina at Chapel Hill, 116 Manning DrivePharmacology & Computational Medicine Program, University of North Carolina at Chapel Hill, 116 Manning DriveDepartment of Biochemistry and Biophysics, University of CaliforniaDepartment of Molecular Biosciences, University of South Florida, 4202 E Fowler AveAbstract Unraveling the signaling roles of intermediate complexes is pivotal for G protein-coupled receptor (GPCR) drug development. Despite hundreds of GPCR-Gαβγ structures, these snapshots primarily capture the fully activated complex. Consequently, the functions of intermediate GPCR-G protein complexes remain elusive. Guided by a conformational landscape visualized via 19F quantitative NMR and molecular dynamics (MD) simulations, we determined the structure of an intermediate GPCR-mini-Gαsβγ complex at 2.6 Å using cryo-EM, by blocking its transition to the fully activated complex. Furthermore, we present direct evidence that the complex at this intermediate state initiates a rate-limited nucleotide exchange before transitioning to the fully activated complex. In this state, BODIPY-GDP/GTP based nucleotide exchange assays further indicated the α-helical domain of the Gα is partially open, allowing it to grasp a nucleotide at a non-canonical binding site, distinct from the canonical nucleotide-binding site. These advances bridge a significant gap in our understanding of the complexity of GPCR signaling.https://doi.org/10.1038/s41467-025-56434-4 |
| spellingShingle | Maxine Bi Xudong Wang Jinan Wang Jun Xu Wenkai Sun Victor Ayo Adediwura Yinglong Miao Yifan Cheng Libin Ye Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex Nature Communications |
| title | Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex |
| title_full | Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex |
| title_fullStr | Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex |
| title_full_unstemmed | Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex |
| title_short | Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex |
| title_sort | structure and function of a near fully activated intermediate gpcr gαβγ complex |
| url | https://doi.org/10.1038/s41467-025-56434-4 |
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