Cryo-EM structures reveal native GABAA receptor assemblies and pharmacology

Sun, Chang; Zhu, Hongtao; Clark, Sarah; Gouaux, Eric

doi:10.1038/s41586-023-06556-w

Cryo-EM structures reveal native GABA_A receptor assemblies and pharmacology

Journal Article · Wed Sep 20 00:00:00 EDT 2023 · Nature (London)

DOI:https://doi.org/10.1038/s41586-023-06556-w· OSTI ID:2471665

Sun, Chang ^[1]; Zhu, Hongtao ^[1]; Clark, Sarah ^[1]; ^[1]

Oregon Health & Science Univ., Portland, OR (United States)

Type A γ-aminobutyric acid receptors (GABA_ARs) are the principal inhibitory receptors in the brain and the target of a wide range of clinical agents, including anaesthetics, sedatives, hypnotics and antidepressants. However, our understanding of GABA_AR pharmacology has been hindered by the vast number of pentameric assemblies that can be derived from 19 different subunits and the lack of structural knowledge of clinically relevant receptors. Here, we isolate native murine GABA_AR assemblies containing the widely expressed α1 subunit and elucidate their structures in complex with drugs used to treat insomnia (zolpidem (ZOL) and flurazepam) and postpartum depression (the neurosteroid allopregnanolone (APG)). Using cryo-electron microscopy (cryo-EM) analysis and single-molecule photobleaching experiments, we uncover three major structural populations in the brain: the canonical α1β2γ2 receptor containing two α1 subunits, and two assemblies containing one α1 and either an α2 or α3 subunit, in which the single α1-containing receptors feature a more compact arrangement between the transmembrane and extracellular domains. Interestingly, APG is bound at the transmembrane α/β subunit interface, even when not added to the sample, revealing an important role for endogenous neurosteroids in modulating native GABA_ARs. Together with structurally engaged lipids, neurosteroids produce global conformational changes throughout the receptor that modify the ion channel pore and the binding sites for GABA and insomnia medications. Our data reveal the major α1-containing GABA_AR assemblies, bound with endogenous neurosteroid, thus defining a structural landscape from which subtype-specific drugs can be developed.

View Accepted Manuscript (DOE)

Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 2471665

Journal Information:: Nature (London), Journal Name: Nature (London) Journal Issue: 7981 Vol. 622; ISSN 0028-0836

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Cryo-EM structures reveal native GABAA receptor assemblies and pharmacology

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