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A neurodevelopmental disorder mutation locks G proteins in the transitory pre-activated state

Author

Listed:
  • Kevin M. Knight

    (University of North Carolina at Chapel Hill
    University of Florida)

  • Brian E. Krumm

    (University of North Carolina at Chapel Hill)

  • Nicholas J. Kapolka

    (University of North Carolina at Chapel Hill)

  • W. Grant Ludlam

    (University of Florida)

  • Meng Cui

    (Department of Pharmaceutical Sciences Northeastern University)

  • Sepehr Mani

    (Department of Pharmaceutical Sciences Northeastern University)

  • Iya Prytkova

    (Icahn School of Medicine at Mount Sinai)

  • Elizabeth G. Obarow

    (University of North Carolina at Chapel Hill)

  • Tyler J. Lefevre

    (University of Michigan)

  • Wenyuan Wei

    (Beckman Research Institute of the City of Hope)

  • Ning Ma

    (Beckman Research Institute of the City of Hope)

  • Xi-Ping Huang

    (University of North Carolina at Chapel Hill)

  • Jonathan F. Fay

    (Baltimore)

  • Nagarajan Vaidehi

    (Beckman Research Institute of the City of Hope)

  • Alan V. Smrcka

    (University of Michigan)

  • Paul A. Slesinger

    (Icahn School of Medicine at Mount Sinai)

  • Diomedes E. Logothetis

    (Department of Pharmaceutical Sciences Northeastern University)

  • Kirill A. Martemyanov

    (University of Florida)

  • Bryan L. Roth

    (University of North Carolina at Chapel Hill)

  • Henrik G. Dohlman

    (University of North Carolina at Chapel Hill)

Abstract

Many neurotransmitter receptors activate G proteins through exchange of GDP for GTP. The intermediate nucleotide-free state has eluded characterization, due largely to its inherent instability. Here we characterize a G protein variant associated with a rare neurological disorder in humans. GαoK46E has a charge reversal that clashes with the phosphate groups of GDP and GTP. As anticipated, the purified protein binds poorly to guanine nucleotides yet retains wild-type affinity for G protein βγ subunits. In cells with physiological concentrations of nucleotide, GαoK46E forms a stable complex with receptors and Gβγ, impeding effector activation. Further, we demonstrate that the mutant can be easily purified in complex with dopamine-bound D2 receptors, and use cryo-electron microscopy to determine the structure, including both domains of Gαo, without nucleotide or stabilizing nanobodies. These findings reveal the molecular basis for the first committed step of G protein activation, establish a mechanistic basis for a neurological disorder, provide a simplified strategy to determine receptor-G protein structures, and a method to detect high affinity agonist binding in cells.

Suggested Citation

  • Kevin M. Knight & Brian E. Krumm & Nicholas J. Kapolka & W. Grant Ludlam & Meng Cui & Sepehr Mani & Iya Prytkova & Elizabeth G. Obarow & Tyler J. Lefevre & Wenyuan Wei & Ning Ma & Xi-Ping Huang & Jona, 2024. "A neurodevelopmental disorder mutation locks G proteins in the transitory pre-activated state," Nature Communications, Nature, vol. 15(1), pages 1-18, December.
  • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-50964-z
    DOI: 10.1038/s41467-024-50964-z
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    References listed on IDEAS

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    1. Tilman Flock & Alexander S. Hauser & Nadia Lund & David E. Gloriam & Santhanam Balaji & M. Madan Babu, 2017. "Selectivity determinants of GPCR–G-protein binding," Nature, Nature, vol. 545(7654), pages 317-322, May.
    2. Tristan Bepler & Kotaro Kelley & Alex J. Noble & Bonnie Berger, 2020. "Topaz-Denoise: general deep denoising models for cryoEM and cryoET," Nature Communications, Nature, vol. 11(1), pages 1-12, December.
    3. Christopher J. Draper-Joyce & Maryam Khoshouei & David M. Thal & Yi-Lynn Liang & Anh T. N. Nguyen & Sebastian G. B. Furness & Hariprasad Venugopal & Jo-Anne Baltos & Jürgen M. Plitzko & Radostin Danev, 2018. "Structure of the adenosine-bound human adenosine A1 receptor–Gi complex," Nature, Nature, vol. 558(7711), pages 559-563, June.
    4. Yi-Lynn Liang & Maryam Khoshouei & Alisa Glukhova & Sebastian G. B. Furness & Peishen Zhao & Lachlan Clydesdale & Cassandra Koole & Tin T. Truong & David M. Thal & Saifei Lei & Mazdak Radjainia & Rado, 2018. "Phase-plate cryo-EM structure of a biased agonist-bound human GLP-1 receptor–Gs complex," Nature, Nature, vol. 555(7694), pages 121-125, March.
    5. Brian T. DeVree & Jacob P. Mahoney & Gisselle A. Vélez-Ruiz & Soren G. F. Rasmussen & Adam J. Kuszak & Elin Edwald & Juan-Jose Fung & Aashish Manglik & Matthieu Masureel & Yang Du & Rachel A. Matt & E, 2016. "Allosteric coupling from G protein to the agonist-binding pocket in GPCRs," Nature, Nature, vol. 535(7610), pages 182-186, July.
    6. Ka Young Chung & Søren G. F. Rasmussen & Tong Liu & Sheng Li & Brian T. DeVree & Pil Seok Chae & Diane Calinski & Brian K. Kobilka & Virgil L. Woods & Roger K. Sunahara, 2011. "Conformational changes in the G protein Gs induced by the β2 adrenergic receptor," Nature, Nature, vol. 477(7366), pages 611-615, September.
    7. Weijiao Huang & Aashish Manglik & A. J. Venkatakrishnan & Toon Laeremans & Evan N. Feinberg & Adrian L. Sanborn & Hideaki E. Kato & Kathryn E. Livingston & Thor S. Thorsen & Ralf C. Kling & Sébastien , 2015. "Structural insights into µ-opioid receptor activation," Nature, Nature, vol. 524(7565), pages 315-321, August.
    8. Ramona Schrage & Anna-Lena Schmitz & Evelyn Gaffal & Suvi Annala & Stefan Kehraus & Daniela Wenzel & Katrin M. Büllesbach & Tobias Bald & Asuka Inoue & Yuji Shinjo & Ségolène Galandrin & Naveen Shridh, 2015. "The experimental power of FR900359 to study Gq-regulated biological processes," Nature Communications, Nature, vol. 6(1), pages 1-17, December.
    9. Sheng Wang & Tao Che & Anat Levit & Brian K. Shoichet & Daniel Wacker & Bryan L. Roth, 2018. "Structure of the D2 dopamine receptor bound to the atypical antipsychotic drug risperidone," Nature, Nature, vol. 555(7695), pages 269-273, March.
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