气候变化领域集成服务门户(CCPortal): Modulation of α1β3γ2 GABAA receptors expressed in X. laevis oocytes using a propofol photoswitch tethered to the transmembrane helix

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DOI	10.1073/pnas.2008178118
	Modulation of α1β3γ2 GABAA receptors expressed in X. laevis oocytes using a propofol photoswitch tethered to the transmembrane helix
	Borghese C.M.; Wang H.-Y.L.; McHardy S.F.; Messing R.O.; Trudell J.R.; Harris R.A.; Bertaccini E.J.
发表日期	2021
ISSN	00278424
卷号	118 期号:8
英文摘要	Tethered photoswitches are molecules with two photo-dependent isomeric forms, each with different actions on their biological targets. They include reactive chemical groups capable of covalently binding to their target. Our aim was to develop a β-subunit-tethered propofol photoswitch (MAP20), as a tool to better study the mechanism of anesthesia through the GABAA α1β3γ2 receptor. We used short spacers between the tether (methanethiosulfonate), the photosensitive moiety (azobenzene), and the ligand (propofol), to allow a precise tethering adjacent to the putative propofol binding site at the β+α− interface of the receptor transmembrane helices (TMs). First, we used molecular modeling to identify possible tethering sites in β3TM3 and α1TM1, and then introduced cysteines in the candidate positions. Two mutant subunits [β3(M283C) and α1(V227C)] showed photomodulation of GABA responses after incubation with MAP20 and illumination with lights at specific wavelengths. The α1β3(M283C)γ2 receptor showed the greatest photomodulation, which decreased as GABA concentration increased. The location of the mutations that produced photomodulation confirmed that the propofol binding site is located in the β+α− interface close to the extracellular side of the transmembrane helices. Tethering the photoswitch to cysteines introduced in the positions homologous to β3M283 in two other subunits (α1W288 and γ2L298) also produced photomodulation, which was not entirely reversible, probably reflecting the different nature of each interface. The results are in agreement with a binding site in the β+α− interface for the anesthetic propofol. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Anesthetic; Azobenzene; Methanethiosulfonate; Molecular modeling; Optopharmcology
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180557
作者单位	Waggoner Center for Alcohol and Addiction Research, University of Texas at Austin, Austin, TX 78712, United States; Department of Neuroscience, University of Texas at Austin, Austin, TX 78712, United States; Center for Innovative Drug Discovery, University of Texas at San Antonio, San Antonio, TX 78249, United States; Department of Anesthesia, Stanford University, Palo Alto, CA 94305, United States; Beckman Program for Molecular and Genetic Medicine, Stanford University, Palo Alto, CA 94305, United States; Department of Anesthesia, Palo Alto VA Health Care System, Palo Alto Division, Palo Alto, CA 94304, United States
推荐引用方式 GB/T 7714	Borghese C.M.,Wang H.-Y.L.,McHardy S.F.,et al. Modulation of α1β3γ2 GABAA receptors expressed in X. laevis oocytes using a propofol photoswitch tethered to the transmembrane helix[J],2021,118(8).
APA	Borghese C.M..,Wang H.-Y.L..,McHardy S.F..,Messing R.O..,Trudell J.R..,...&Bertaccini E.J..(2021).Modulation of α1β3γ2 GABAA receptors expressed in X. laevis oocytes using a propofol photoswitch tethered to the transmembrane helix.Proceedings of the National Academy of Sciences of the United States of America,118(8).
MLA	Borghese C.M.,et al."Modulation of α1β3γ2 GABAA receptors expressed in X. laevis oocytes using a propofol photoswitch tethered to the transmembrane helix".Proceedings of the National Academy of Sciences of the United States of America 118.8(2021).