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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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