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Characterization of Binding Site Interactions and Selectivity Principles in the α3β4 Nicotinic Acetylcholine Receptor

Knox, Hailey J. and Rego Campello, Hugo and Lester, Henry A. and Gallagher, Timothy and Dougherty, Dennis A. (2022) Characterization of Binding Site Interactions and Selectivity Principles in the α3β4 Nicotinic Acetylcholine Receptor. Journal of the American Chemical Society, 144 (35). pp. 16101-16117. ISSN 0002-7863. doi:10.1021/jacs.2c06495.

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Nicotinic acetylcholine receptors (nAChRs) play an important role in neurotransmission and are also involved in addiction and several disease states. There is significant interest in therapeutic targeting of nAChRs; however, achieving selectivity for one subtype over others has been a longstanding challenge, given the close structural similarities across the family. Here, we characterize binding interactions in the α3β4 nAChR subtype via structure–function studies involving noncanonical amino acid mutagenesis and two-electrode voltage clamp electrophysiology. We establish comprehensive binding models for both the endogenous neurotransmitter ACh and the smoking cessation drug cytisine. We also use a panel of C(10)-substituted cytisine derivatives to probe the effects of subtle changes in the ligand structure on binding. By comparing our results to those obtained for the well-studied α4β2 subtype, we identify several features of both the receptor and agonist structure that can be utilized to enhance selectivity for either α3β4 or α4β2. Finally, we characterize binding interactions of the α3β4-selective partial agonist AT-1001 to determine factors that contribute to its selectivity. These results shed new light on the design of selective nAChR-targeted ligands and can be used to inform the design of improved therapies with minimized off-target effects.

Item Type:Article
Related URLs:
URLURL TypeDescription
Knox, Hailey J.0000-0003-0608-2855
Rego Campello, Hugo0000-0001-8588-0198
Lester, Henry A.0000-0002-5470-5255
Gallagher, Timothy0000-0002-3544-327X
Dougherty, Dennis A.0000-0003-1464-2461
Additional Information:We would like to thank Jonathan Wang and Purnima Deshpande for harvesting and preparing oocytes from Xenopus laevis. We thank Stephen Grant, Maria Constanza Maldifassi Gatica, and Christopher Marotta for useful discussions and suggestions. We thank Annet Blom for guidance on initial expression and stoichiometry determination for α3β4. We also acknowledge Achieve Life Sciences for their generous gift of (−)-cytisine. This work was supported by in part by funds provided by The Regents of the University of California, Research Grants Program Office, Tobacco Related Disease Research Program (Grant No. T29IR0455 to D.A.D.). The opinions, findings, and conclusions herein are those of the authors and do not necessarily represent those of The Regents of the University of California, or any of its programs.
Funding AgencyGrant Number
California Tobacco-Related Disease Research ProgramT29IR0455
Issue or Number:35
Record Number:CaltechAUTHORS:20220909-232673000
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:116865
Deposited By: Donna Wrublewski
Deposited On:03 Dec 2022 00:31
Last Modified:03 Dec 2022 00:31

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