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Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex

Author

Listed:
  • Maxine Bi

    (University of California)

  • Xudong Wang

    (4202 E Fowler Ave)

  • Jinan Wang

    (University of North Carolina at Chapel Hill, 116 Manning Drive)

  • Jun Xu

    (Stanford University School of Medicine)

  • Wenkai Sun

    (4202 E Fowler Ave)

  • Victor Ayo Adediwura

    (University of North Carolina at Chapel Hill, 116 Manning Drive)

  • Yinglong Miao

    (University of North Carolina at Chapel Hill, 116 Manning Drive)

  • Yifan Cheng

    (University of California
    University of California)

  • Libin Ye

    (4202 E Fowler Ave
    12902 USF Magnolia Drive)

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.

Suggested Citation

  • Maxine Bi & Xudong Wang & Jinan Wang & Jun Xu & Wenkai Sun & Victor Ayo Adediwura & Yinglong Miao & Yifan Cheng & Libin Ye, 2025. "Structure and function of a near fully-activated intermediate GPCR-Gαβγ complex," Nature Communications, Nature, vol. 16(1), pages 1-14, December.
  • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-56434-4
    DOI: 10.1038/s41467-025-56434-4
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    References listed on IDEAS

    as
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