@article{0a6b8d76d5694416906b1b1fc8b257a2,
title = "Agonist Selectivity and Ion Permeation in the α3β4 Ganglionic Nicotinic Receptor",
abstract = "Nicotinic acetylcholine receptors are pentameric ion channels that mediate fast chemical neurotransmission. The α3β4 nicotinic receptor subtype forms the principal relay between the central and peripheral nervous systems in the autonomic ganglia. This receptor is also expressed focally in brain areas that affect reward circuits and addiction. Here, we present structures of the α3β4 nicotinic receptor in lipidic and detergent environments, using functional reconstitution to define lipids appropriate for structural analysis. The structures of the receptor in complex with nicotine, as well as the α3β4-selective ligand AT-1001, complemented by molecular dynamics, suggest principles of agonist selectivity. The structures further reveal much of the architecture of the intracellular domain, where mutagenesis experiments and simulations define residues governing ion conductance.",
keywords = "addiction, cryo-EM, ganglionic, ligand-gated ion channel, lipids, nicotine, nicotinic receptor",
author = "Anant Gharpure and Jinfeng Teng and Yuxuan Zhuang and Noviello, {Colleen M.} and Walsh, {Richard M.} and Rico Cabuco and Howard, {Rebecca J.} and Zaveri, {Nurulain T.} and Erik Lindahl and Hibbs, {Ryan E.}",
note = "Funding Information: We thank D. Cawley for antibody production, X. Bai for EM discussion, P. Blount, R. Syeda, and W. Zeng for liposome patch advice, J. Frauenfeld (Salipro Biotech) for the saposin expression construct, and all members of the Hibbs lab for discussion. Single-particle cryo-EM data were collected at the University of Texas Southwestern Medical Center Cryo-Electron Microscopy Facility, which is supported by the CPRIT Core Facility Support Award RP170644. A.G. acknowledges support from the NIH (T32DA07290). R.W. acknowledges support from the Sara and Frank McKnight Fund for Biochemical Research and the NIH (T32GM008203). This work was supported by a McKnight Scholar Award, The Welch Foundation (I-1812), and grants from the NIH (DA037492, DA042072, and NS095899) to R.E.H. A.G. and R.E.H. conceived the project. A.G. performed the protein production, purification, structural analysis, binding experiments, and cell-attached electrophysiology. J.T. performed the whole-cell and liposome patch-clamp experiments. C.M.N. and R.W. collected the EM data. Y.Z. R.J.H. and E.L. performed molecular dynamics simulations. R.C. assisted with baculovirus production. N.T.Z. provided AT-1001. A.G. and R.E.H. wrote the manuscript with input from all other authors. N.T.Z. is the founder and an employee of Astraea Therapeutics and a member of its scientific advisory board. Funding Information: We thank D. Cawley for antibody production, X. Bai for EM discussion, P. Blount, R. Syeda, and W. Zeng for liposome patch advice, J. Frauenfeld (Salipro Biotech) for the saposin expression construct, and all members of the Hibbs lab for discussion. Single-particle cryo-EM data were collected at the University of Texas Southwestern Medical Center Cryo-Electron Microscopy Facility, which is supported by the CPRIT Core Facility Support Award RP170644 . A.G. acknowledges support from the NIH ( T32DA07290 ). R.W. acknowledges support from the Sara and Frank McKnight Fund for Biochemical Research and the NIH ( T32GM008203 ). This work was supported by a McKnight Scholar Award , The Welch Foundation ( I-1812 ), and grants from the NIH ( DA037492 , DA042072 , and NS095899 ) to R.E.H. Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
month = nov,
day = "6",
doi = "10.1016/j.neuron.2019.07.030",
language = "English (US)",
volume = "104",
pages = "501--511.e6",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "3",
}