Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD

Can Cao; Ximena Barros-Álvarez; Shicheng Zhang; Kuglae Kim; Marc A. Dämgen; Ouliana Panova; Carl Mikael Suomivuori; Jonathan F. Fay; Xiaofang Zhong; Brian E. Krumm; Ryan H. Gumpper; Alpay B. Seven; Michael J. Robertson; Nevan J. Krogan; Ruth Hüttenhain; David E. Nichols; Ron O. Dror; Georgios Skiniotis; Bryan L. Roth

doi:10.1016/j.neuron.2022.08.006

Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD

Can Cao, Ximena Barros-Álvarez, Shicheng Zhang, Kuglae Kim, Marc A. Dämgen, Ouliana Panova, Carl Mikael Suomivuori, Jonathan F. Fay, Xiaofang Zhong, Brian E. Krumm, Ryan H. Gumpper, Alpay B. Seven, Michael J. Robertson, Nevan J. Krogan, Ruth Hüttenhain, David E. Nichols, Ron O. Dror, Georgios Skiniotis, Bryan L. Roth

Department of Pharmacy

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

Serotonin (5-hydroxytryptamine [5-HT]) 5-HT2-family receptors represent essential targets for lysergic acid diethylamide (LSD) and all other psychedelic drugs. Although the primary psychedelic drug effects are mediated by the 5-HT_2A serotonin receptor (HTR2A), the 5-HT_2B serotonin receptor (HTR2B) has been used as a model receptor to study the activation mechanisms of psychedelic drugs due to its high expression and similarity to HTR2A. In this study, we determined the cryo-EM structures of LSD-bound HTR2B in the transducer-free, Gq-protein-coupled, and β-arrestin-1-coupled states. These structures provide distinct signaling snapshots of LSD's action, ranging from the transducer-free, partially active state to the transducer-coupled, fully active states. Insights from this study will both provide comprehensive molecular insights into the signaling mechanisms of the prototypical psychedelic LSD and accelerate the discovery of novel psychedelic drugs.

Original language	English
Pages (from-to)	3154-3167.e7
Journal	Neuron
Volume	110
Issue number	19
DOIs	https://doi.org/10.1016/j.neuron.2022.08.006
Publication status	Published - 2022 Oct 5

Bibliographical note

Funding Information:
We thank Elizabeth Montabana at the Stanford cEMc facility for support with data collection. M.A.D. was supported by a Walter Benjamin Fellowship from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) project number 471315262. C.-M.S. is supported by a Long-Term Fellowship (LT000916/2018-L) from the Human Frontier Science Program (HFSP). R.O.D. G.S. and B.L.R. are supported by the Defense Advanced Research Projects Agency (DARPA) under HR0011-20-2-0029. N.J.K. and R.H. are supported by the Defense Advanced Research Projects Agency (DARPA) under the Cooperative Agreements HR0011-19-2-0020 and HR0011-20-2-0029. 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. An award for computer time was provided by the INCITE program. This research used resources of the Oak Ridge Leadership Computing Facility, which is a DOE Office of Science User Facility supported under contract DE-AC05-00OR22725. B.L.R. and others in the Roth lab were also supported by the Michael Hooker Distinguished Professorship and NIH grants R37DA045657 and RO1MH112205. This work was also supported by NIH grant R01GM127359 (R.O.D.). C.C. initiated the project, performed the initial arrestin recruitment assay, established the whole protocol for arresting complex formation, performed the cloning, expression, and purification of the HTR2B-β-arrestin-1-scFv30 complex and transducer-free HTR2B, analyzed the structures, and wrote the manuscript. X.B.-A. evaluated the protein samples, prepared the cryo-EM grids, oversaw the data collection, processed the cryo-EM data, modeled the structures, analyzed the structural data, and prepared the data-processing figures. S.Z. helped with the protein expression, performed the BRET1 assays, and analyzed the functional data. K.K. purified the HTR2B-Gq complex. M.A.D. and C.-M.S. performed and analyzed the MD simulations. O.P. prepared the cryo-EM grids and performed the cryo-EM data collection. B.E.K. and R.H.G. reviewed the manuscript. A.B.S. assisted in the cryo-EM data processing. M.J.R. assisted with the ligand pose validations. X.Z. N.J.K. and R.H. performed the mass spectrometry analysis of HTR2B C-tail. D.E.N. provided insight into LSD action. J.F.F. performed the initial cryo-EM analysis for the HTR2B-β-arrestin-1-scFv30 sample. R.O.D. supervised the MD simulation studies and edited the manuscript. G.S. supervised the cryo-EM studies and edited the manuscript. B.L.R. supervised the entire project and edited the manuscript. The Krogan Laboratory has received research support from Vir Biotechnology, F. Hoffmann-La Roche, and Rezo Therapeutics. N.J.K. has financially compensated consulting agreements with the Icahn School of Medicine at Mount Sinai, New York; Maze Therapeutics; Interline Therapeutics; Rezo Therapeutics; GEn1E Life Sciences, Inc.; and Twist Bioscience Corp. N.J.K. is on the Board of Directors of Rezo Therapeutics and is a shareholder in Tenaya Therapeutics, Maze Therapeutics, Rezo Therapeutics, and Interline Therapeutics. G.S. is co-founder and consultant for Deep Apple Therapeutics.

Funding Information:
We thank Elizabeth Montabana at the Stanford cEMc facility for support with data collection. M.A.D. was supported by a Walter Benjamin Fellowship from the Deutsche Forschungsgemeinschaft ( DFG , German Research Foundation) project number 471315262 . C.-M.S. is supported by a Long-Term Fellowship ( LT000916/2018-L ) from the Human Frontier Science Program ( HFSP ). R.O.D., G.S., and B.L.R. are supported by the Defense Advanced Research Projects Agency ( DARPA ) under HR0011-20-2-0029 . N.J.K. and R.H. are supported by the Defense Advanced Research Projects Agency (DARPA) under the Cooperative Agreements HR0011-19-2-0020 and HR0011-20-2-0029 . 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. An award for computer time was provided by the INCITE program. This research used resources of the Oak Ridge Leadership Computing Facility, which is a DOE Office of Science User Facility supported under contract DE-AC05-00OR22725. B.L.R. and others in the Roth lab were also supported by the Michael Hooker Distinguished Professorship and NIH grants R37DA045657 and RO1MH112205 . This work was also supported by NIH grant R01GM127359 (R.O.D.).

Publisher Copyright:
© 2022 Elsevier Inc.

All Science Journal Classification (ASJC) codes

Neuroscience(all)

Access to Document

10.1016/j.neuron.2022.08.006

Cite this

Cao, C., Barros-Álvarez, X., Zhang, S., Kim, K., Dämgen, M. A., Panova, O., Suomivuori, C. M., Fay, J. F., Zhong, X., Krumm, B. E., Gumpper, R. H., Seven, A. B., Robertson, M. J., Krogan, N. J., Hüttenhain, R., Nichols, D. E., Dror, R. O., Skiniotis, G., & Roth, B. L. (2022). Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD. Neuron, 110(19), 3154-3167.e7. https://doi.org/10.1016/j.neuron.2022.08.006

@article{8b109dd63de04fd3bb020f27700acd3e,

title = "Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD",

abstract = "Serotonin (5-hydroxytryptamine [5-HT]) 5-HT2-family receptors represent essential targets for lysergic acid diethylamide (LSD) and all other psychedelic drugs. Although the primary psychedelic drug effects are mediated by the 5-HT2A serotonin receptor (HTR2A), the 5-HT2B serotonin receptor (HTR2B) has been used as a model receptor to study the activation mechanisms of psychedelic drugs due to its high expression and similarity to HTR2A. In this study, we determined the cryo-EM structures of LSD-bound HTR2B in the transducer-free, Gq-protein-coupled, and β-arrestin-1-coupled states. These structures provide distinct signaling snapshots of LSD's action, ranging from the transducer-free, partially active state to the transducer-coupled, fully active states. Insights from this study will both provide comprehensive molecular insights into the signaling mechanisms of the prototypical psychedelic LSD and accelerate the discovery of novel psychedelic drugs.",

author = "Can Cao and Ximena Barros-{\'A}lvarez and Shicheng Zhang and Kuglae Kim and D{\"a}mgen, {Marc A.} and Ouliana Panova and Suomivuori, {Carl Mikael} and Fay, {Jonathan F.} and Xiaofang Zhong and Krumm, {Brian E.} and Gumpper, {Ryan H.} and Seven, {Alpay B.} and Robertson, {Michael J.} and Krogan, {Nevan J.} and Ruth H{\"u}ttenhain and Nichols, {David E.} and Dror, {Ron O.} and Georgios Skiniotis and Roth, {Bryan L.}",

note = "Funding Information: We thank Elizabeth Montabana at the Stanford cEMc facility for support with data collection. M.A.D. was supported by a Walter Benjamin Fellowship from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) project number 471315262. C.-M.S. is supported by a Long-Term Fellowship (LT000916/2018-L) from the Human Frontier Science Program (HFSP). R.O.D. G.S. and B.L.R. are supported by the Defense Advanced Research Projects Agency (DARPA) under HR0011-20-2-0029. N.J.K. and R.H. are supported by the Defense Advanced Research Projects Agency (DARPA) under the Cooperative Agreements HR0011-19-2-0020 and HR0011-20-2-0029. 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. An award for computer time was provided by the INCITE program. This research used resources of the Oak Ridge Leadership Computing Facility, which is a DOE Office of Science User Facility supported under contract DE-AC05-00OR22725. B.L.R. and others in the Roth lab were also supported by the Michael Hooker Distinguished Professorship and NIH grants R37DA045657 and RO1MH112205. This work was also supported by NIH grant R01GM127359 (R.O.D.). C.C. initiated the project, performed the initial arrestin recruitment assay, established the whole protocol for arresting complex formation, performed the cloning, expression, and purification of the HTR2B-β-arrestin-1-scFv30 complex and transducer-free HTR2B, analyzed the structures, and wrote the manuscript. X.B.-A. evaluated the protein samples, prepared the cryo-EM grids, oversaw the data collection, processed the cryo-EM data, modeled the structures, analyzed the structural data, and prepared the data-processing figures. S.Z. helped with the protein expression, performed the BRET1 assays, and analyzed the functional data. K.K. purified the HTR2B-Gq complex. M.A.D. and C.-M.S. performed and analyzed the MD simulations. O.P. prepared the cryo-EM grids and performed the cryo-EM data collection. B.E.K. and R.H.G. reviewed the manuscript. A.B.S. assisted in the cryo-EM data processing. M.J.R. assisted with the ligand pose validations. X.Z. N.J.K. and R.H. performed the mass spectrometry analysis of HTR2B C-tail. D.E.N. provided insight into LSD action. J.F.F. performed the initial cryo-EM analysis for the HTR2B-β-arrestin-1-scFv30 sample. R.O.D. supervised the MD simulation studies and edited the manuscript. G.S. supervised the cryo-EM studies and edited the manuscript. B.L.R. supervised the entire project and edited the manuscript. The Krogan Laboratory has received research support from Vir Biotechnology, F. Hoffmann-La Roche, and Rezo Therapeutics. N.J.K. has financially compensated consulting agreements with the Icahn School of Medicine at Mount Sinai, New York; Maze Therapeutics; Interline Therapeutics; Rezo Therapeutics; GEn1E Life Sciences, Inc.; and Twist Bioscience Corp. N.J.K. is on the Board of Directors of Rezo Therapeutics and is a shareholder in Tenaya Therapeutics, Maze Therapeutics, Rezo Therapeutics, and Interline Therapeutics. G.S. is co-founder and consultant for Deep Apple Therapeutics. Funding Information: We thank Elizabeth Montabana at the Stanford cEMc facility for support with data collection. M.A.D. was supported by a Walter Benjamin Fellowship from the Deutsche Forschungsgemeinschaft ( DFG , German Research Foundation) project number 471315262 . C.-M.S. is supported by a Long-Term Fellowship ( LT000916/2018-L ) from the Human Frontier Science Program ( HFSP ). R.O.D., G.S., and B.L.R. are supported by the Defense Advanced Research Projects Agency ( DARPA ) under HR0011-20-2-0029 . N.J.K. and R.H. are supported by the Defense Advanced Research Projects Agency (DARPA) under the Cooperative Agreements HR0011-19-2-0020 and HR0011-20-2-0029 . 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. An award for computer time was provided by the INCITE program. This research used resources of the Oak Ridge Leadership Computing Facility, which is a DOE Office of Science User Facility supported under contract DE-AC05-00OR22725. B.L.R. and others in the Roth lab were also supported by the Michael Hooker Distinguished Professorship and NIH grants R37DA045657 and RO1MH112205 . This work was also supported by NIH grant R01GM127359 (R.O.D.). Publisher Copyright: {\textcopyright} 2022 Elsevier Inc.",

year = "2022",

month = oct,

day = "5",

doi = "10.1016/j.neuron.2022.08.006",

language = "English",

volume = "110",

pages = "3154--3167.e7",

journal = "Neuron",

issn = "0896-6273",

publisher = "Cell Press",

number = "19",

}

Cao, C, Barros-Álvarez, X, Zhang, S, Kim, K, Dämgen, MA, Panova, O, Suomivuori, CM, Fay, JF, Zhong, X, Krumm, BE, Gumpper, RH, Seven, AB, Robertson, MJ, Krogan, NJ, Hüttenhain, R, Nichols, DE, Dror, RO, Skiniotis, G & Roth, BL 2022, 'Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD', Neuron, vol. 110, no. 19, pp. 3154-3167.e7. https://doi.org/10.1016/j.neuron.2022.08.006

TY - JOUR

T1 - Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD

AU - Cao, Can

AU - Barros-Álvarez, Ximena

AU - Zhang, Shicheng

AU - Kim, Kuglae

AU - Dämgen, Marc A.

AU - Panova, Ouliana

AU - Suomivuori, Carl Mikael

AU - Fay, Jonathan F.

AU - Zhong, Xiaofang

AU - Krumm, Brian E.

AU - Gumpper, Ryan H.

AU - Seven, Alpay B.

AU - Robertson, Michael J.

AU - Krogan, Nevan J.

AU - Hüttenhain, Ruth

AU - Nichols, David E.

AU - Dror, Ron O.

AU - Skiniotis, Georgios

AU - Roth, Bryan L.

N1 - Funding Information: We thank Elizabeth Montabana at the Stanford cEMc facility for support with data collection. M.A.D. was supported by a Walter Benjamin Fellowship from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) project number 471315262. C.-M.S. is supported by a Long-Term Fellowship (LT000916/2018-L) from the Human Frontier Science Program (HFSP). R.O.D. G.S. and B.L.R. are supported by the Defense Advanced Research Projects Agency (DARPA) under HR0011-20-2-0029. N.J.K. and R.H. are supported by the Defense Advanced Research Projects Agency (DARPA) under the Cooperative Agreements HR0011-19-2-0020 and HR0011-20-2-0029. 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. An award for computer time was provided by the INCITE program. This research used resources of the Oak Ridge Leadership Computing Facility, which is a DOE Office of Science User Facility supported under contract DE-AC05-00OR22725. B.L.R. and others in the Roth lab were also supported by the Michael Hooker Distinguished Professorship and NIH grants R37DA045657 and RO1MH112205. This work was also supported by NIH grant R01GM127359 (R.O.D.). C.C. initiated the project, performed the initial arrestin recruitment assay, established the whole protocol for arresting complex formation, performed the cloning, expression, and purification of the HTR2B-β-arrestin-1-scFv30 complex and transducer-free HTR2B, analyzed the structures, and wrote the manuscript. X.B.-A. evaluated the protein samples, prepared the cryo-EM grids, oversaw the data collection, processed the cryo-EM data, modeled the structures, analyzed the structural data, and prepared the data-processing figures. S.Z. helped with the protein expression, performed the BRET1 assays, and analyzed the functional data. K.K. purified the HTR2B-Gq complex. M.A.D. and C.-M.S. performed and analyzed the MD simulations. O.P. prepared the cryo-EM grids and performed the cryo-EM data collection. B.E.K. and R.H.G. reviewed the manuscript. A.B.S. assisted in the cryo-EM data processing. M.J.R. assisted with the ligand pose validations. X.Z. N.J.K. and R.H. performed the mass spectrometry analysis of HTR2B C-tail. D.E.N. provided insight into LSD action. J.F.F. performed the initial cryo-EM analysis for the HTR2B-β-arrestin-1-scFv30 sample. R.O.D. supervised the MD simulation studies and edited the manuscript. G.S. supervised the cryo-EM studies and edited the manuscript. B.L.R. supervised the entire project and edited the manuscript. The Krogan Laboratory has received research support from Vir Biotechnology, F. Hoffmann-La Roche, and Rezo Therapeutics. N.J.K. has financially compensated consulting agreements with the Icahn School of Medicine at Mount Sinai, New York; Maze Therapeutics; Interline Therapeutics; Rezo Therapeutics; GEn1E Life Sciences, Inc.; and Twist Bioscience Corp. N.J.K. is on the Board of Directors of Rezo Therapeutics and is a shareholder in Tenaya Therapeutics, Maze Therapeutics, Rezo Therapeutics, and Interline Therapeutics. G.S. is co-founder and consultant for Deep Apple Therapeutics. Funding Information: We thank Elizabeth Montabana at the Stanford cEMc facility for support with data collection. M.A.D. was supported by a Walter Benjamin Fellowship from the Deutsche Forschungsgemeinschaft ( DFG , German Research Foundation) project number 471315262 . C.-M.S. is supported by a Long-Term Fellowship ( LT000916/2018-L ) from the Human Frontier Science Program ( HFSP ). R.O.D., G.S., and B.L.R. are supported by the Defense Advanced Research Projects Agency ( DARPA ) under HR0011-20-2-0029 . N.J.K. and R.H. are supported by the Defense Advanced Research Projects Agency (DARPA) under the Cooperative Agreements HR0011-19-2-0020 and HR0011-20-2-0029 . 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. An award for computer time was provided by the INCITE program. This research used resources of the Oak Ridge Leadership Computing Facility, which is a DOE Office of Science User Facility supported under contract DE-AC05-00OR22725. B.L.R. and others in the Roth lab were also supported by the Michael Hooker Distinguished Professorship and NIH grants R37DA045657 and RO1MH112205 . This work was also supported by NIH grant R01GM127359 (R.O.D.). Publisher Copyright: © 2022 Elsevier Inc.

PY - 2022/10/5

Y1 - 2022/10/5

N2 - Serotonin (5-hydroxytryptamine [5-HT]) 5-HT2-family receptors represent essential targets for lysergic acid diethylamide (LSD) and all other psychedelic drugs. Although the primary psychedelic drug effects are mediated by the 5-HT2A serotonin receptor (HTR2A), the 5-HT2B serotonin receptor (HTR2B) has been used as a model receptor to study the activation mechanisms of psychedelic drugs due to its high expression and similarity to HTR2A. In this study, we determined the cryo-EM structures of LSD-bound HTR2B in the transducer-free, Gq-protein-coupled, and β-arrestin-1-coupled states. These structures provide distinct signaling snapshots of LSD's action, ranging from the transducer-free, partially active state to the transducer-coupled, fully active states. Insights from this study will both provide comprehensive molecular insights into the signaling mechanisms of the prototypical psychedelic LSD and accelerate the discovery of novel psychedelic drugs.

AB - Serotonin (5-hydroxytryptamine [5-HT]) 5-HT2-family receptors represent essential targets for lysergic acid diethylamide (LSD) and all other psychedelic drugs. Although the primary psychedelic drug effects are mediated by the 5-HT2A serotonin receptor (HTR2A), the 5-HT2B serotonin receptor (HTR2B) has been used as a model receptor to study the activation mechanisms of psychedelic drugs due to its high expression and similarity to HTR2A. In this study, we determined the cryo-EM structures of LSD-bound HTR2B in the transducer-free, Gq-protein-coupled, and β-arrestin-1-coupled states. These structures provide distinct signaling snapshots of LSD's action, ranging from the transducer-free, partially active state to the transducer-coupled, fully active states. Insights from this study will both provide comprehensive molecular insights into the signaling mechanisms of the prototypical psychedelic LSD and accelerate the discovery of novel psychedelic drugs.

UR - http://www.scopus.com/inward/record.url?scp=85139008292&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85139008292&partnerID=8YFLogxK

U2 - 10.1016/j.neuron.2022.08.006

DO - 10.1016/j.neuron.2022.08.006

M3 - Article

C2 - 36087581

AN - SCOPUS:85139008292

SN - 0896-6273

VL - 110

SP - 3154-3167.e7

JO - Neuron

JF - Neuron

IS - 19

ER -

Signaling snapshots of a serotonin receptor activated by the prototypical psychedelic LSD

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this