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• W3028664489 endingPage "1064" @default.
• W3028664489 startingPage "1051" @default.
• W3028664489 abstract "Nicotinic acetylcholine receptors (nAChRs) are pentameric ion channels expressed in the central nervous systems. nAChRs containing the α4, β2 and α5 subunits are specifically involved in addictive processes, but their functional architecture is poorly understood due to the intricacy of assembly of these subunits. Here we constrained the subunit assembly by designing fully concatenated human α4β2 and α4β2α5 receptors and characterized their properties by two-electrodes voltage–clamp electrophysiology in Xenopus oocytes. We found that α5-containing nAChRs are irreversibly blocked by methanethiosulfonate (MTS) reagents through a covalent reaction with a cysteine present only in α5. MTS-block experiments establish that the concatemers are expressed in intact form at the oocyte surface, but that reconstitution of nAChRs from loose subunits show inefficient and highly variable assembly of α5 with α4 and β2. Mutational analysis shows that the concatemers assemble both in clockwise and anticlockwise orientations, and that α5 does not contribute to ACh binding from its principal (+) site. Reinvestigation of suspected α5-ligands such as galantamine show no specific effect on α5-containing concatemers. Analysis of the α5-D398N mutation that is linked to smoking and lung cancer shows no significant effect on the electrophysiological function, suggesting that its effect might arise from alteration of other cellular processes. The concatemeric strategy provides a well-characterized platform for mechanistic analysis and screening of human α5-specific ligands." @default.
• W3028664489 created "2020-06-05" @default.
• W3028664489 creator A5024892015 @default.
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• W3028664489 date "2020-05-29" @default.
• W3028664489 modified "2023-10-09" @default.
• W3028664489 title "Concatemers to re-investigate the role of α5 in α4β2 nicotinic receptors" @default.
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• W3028664489 doi "https://doi.org/10.1007/s00018-020-03558-z" @default.
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• W3028664489 hasPublicationYear "2020" @default.
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