Structure of a Hallucinogen-Activated Gq-Coupled 5-HT2A Serotonin Receptor

Kuglae Kim, Tao Che, Ouliana Panova, Jeffrey F. DiBerto, Jiankun Lyu, Brian E. Krumm, Daniel Wacker, Michael J. Robertson, Alpay B. Seven, David E. Nichols, Brian K. Shoichet, Georgios Skiniotis, Bryan L. Roth

Research output: Contribution to journalArticlepeer-review

87 Citations (Scopus)

Abstract

Hallucinogens like lysergic acid diethylamide (LSD), psilocybin, and substituted N-benzyl phenylalkylamines are widely used recreationally with psilocybin being considered as a therapeutic for many neuropsychiatric disorders including depression, anxiety, and substance abuse. How psychedelics mediate their actions—both therapeutic and hallucinogenic—are not understood, although activation of the 5-HT2A serotonin receptor (HTR2A) is key. To gain molecular insights into psychedelic actions, we determined the active-state structure of HTR2A bound to 25-CN-NBOH—a prototypical hallucinogen—in complex with an engineered Gαq heterotrimer by cryoelectron microscopy (cryo-EM). We also obtained the X-ray crystal structures of HTR2A complexed with the arrestin-biased ligand LSD or the inverse agonist methiothepin. Comparisons of these structures reveal determinants responsible for HTR2A-Gαq protein interactions as well as the conformational rearrangements involved in active-state transitions. Given the potential therapeutic actions of hallucinogens, these findings could accelerate the discovery of more selective drugs for the treatment of a variety of neuropsychiatric disorders.

Original languageEnglish
Pages (from-to)1574-1588.e19
JournalCell
Volume182
Issue number6
DOIs
Publication statusPublished - 2020 Sep 17

Bibliographical note

Funding Information:
We thank M.J. Miley and the UNC macromolecular crystallization core for advice and use of their equipment for crystal harvesting and transport, which is supported by the National Cancer Institute ( P30CA016086 ), and the staff of GM/CA@APS, which has been funded by the National Cancer Institute ( ACB-12002 ) and the National Institute of General Medical Sciences ( AGM-12006 ). This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science user facility operated for the DOE Office of Science by Argonne National Laboratory ( DE-AC02-06CH11357 ). K.K. was partially supported by Korea Research Foundation (KRF) postdoctoral fellowship ( 2017R1A6A3A03012547 ).This work was supported by NIH ( R37DA04567 and R01MH112205 ), a cooperative agreement from DARPA ( HR0011-20-2-0029 to B.L.R.), and by NIH ( R35GM122481 to B.K.S.). The views, opinions, and/or findings contained in this material are those of the authors and should not be interpreted as representing the official views, policies, or endorsement of the Department of Defense or the US Government.

Funding Information:
We thank M.J. Miley and the UNC macromolecular crystallization core for advice and use of their equipment for crystal harvesting and transport, which is supported by the National Cancer Institute (P30CA016086), and the staff of GM/CA@APS, which has been funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006). This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science user facility operated for the DOE Office of Science by Argonne National Laboratory (DE-AC02-06CH11357). K.K. was partially supported by Korea Research Foundation (KRF) postdoctoral fellowship (2017R1A6A3A03012547).This work was supported by NIH (R37DA04567 and R01MH112205), a cooperative agreement from DARPA (HR0011-20-2-0029 to B.L.R.), and by NIH (R35GM122481 to B.K.S.). The views, opinions, and/or findings contained in this material are those of the authors and should not be interpreted as representing the official views, policies, or endorsement of the Department of Defense or the US Government. K.K. designed the experiments; performed the cloning, expression, purification, and preparation of the complex, model building and structure refinement in the cryo-EM map; performed the crystallization, data collection, and model building and refinement in the X-ray studies; performed BRET assays; and prepared the manuscript. T.C. designed experiments, performed the binding and functional assays, and prepared the manuscript. O.P. prepared EM grids and collected and processed cryo-EM data with help from A.B.S. J.F.D.B. performed the BRET assays. B.E.K. assisted with the diffraction data collection, processing, and structure validation. B.K.S. supervised the docking studies performed by J.L. M.J.R. assisted with ligand pose validations. D.W. designed crystallization constructs. D.E.N. provided insights into hallucinogen actions and assisted with manuscript preparation. G.S. provided advice for G?q-HTR2A complex preparation, supervised the cryo-EM studies, and assisted with manuscript preparation. B.L.R. was responsible for the overall project strategy and management and prepared the manuscript. The authors declare no competing interests.

Publisher Copyright:
© 2020 Elsevier Inc.

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

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