Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia
Slow-channel congenital myasthenic syndromes (SCCMSs) are rare genetic diseases caused by mutations in muscle nicotinic acetylcholine receptor (nAChR) subunits. Most of the known SCCMS-associated mutations localize at the transmembrane region near the ion pore. Only two SCCMS point mutations are at...
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MDPI AG
2021-02-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/5/1278 |
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| author | Denis Kudryavtsev Anastasia Isaeva Daria Barkova Ekaterina Spirova Renata Mukhutdinova Igor Kasheverov Victor Tsetlin |
| author_facet | Denis Kudryavtsev Anastasia Isaeva Daria Barkova Ekaterina Spirova Renata Mukhutdinova Igor Kasheverov Victor Tsetlin |
| author_sort | Denis Kudryavtsev |
| collection | DOAJ |
| description | Slow-channel congenital myasthenic syndromes (SCCMSs) are rare genetic diseases caused by mutations in muscle nicotinic acetylcholine receptor (nAChR) subunits. Most of the known SCCMS-associated mutations localize at the transmembrane region near the ion pore. Only two SCCMS point mutations are at the extracellular domains near the acetylcholine binding site, α1(G153S) being one of them. In this work, a combination of molecular dynamics, targeted mutagenesis, fluorescent Ca<sup>2+</sup> imaging and patch-clamp electrophysiology has been applied to G153S mutant muscle nAChR to investigate the role of hydrogen bonds formed by Ser 153 with C-loop residues near the acetylcholine-binding site. Introduction of L199T mutation to the C-loop in the vicinity of Ser 153 changed hydrogen bonds distribution, decreased acetylcholine potency (EC<sub>50</sub> 2607 vs. 146 nM) of the double mutant and decay kinetics of acetylcholine-evoked cytoplasmic Ca<sup>2+</sup> rise (τ 14.2 ± 0.3 vs. 34.0 ± 0.4 s). These results shed light on molecular mechanisms of nAChR activation-desensitization and on the involvement of such mechanisms in channelopathy genesis. |
| first_indexed | 2024-03-09T00:29:05Z |
| format | Article |
| id | doaj.art-1883d9d82ce144528da8b6cc88ccab2a |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T00:29:05Z |
| publishDate | 2021-02-01 |
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| spelling | doaj.art-1883d9d82ce144528da8b6cc88ccab2a2023-12-11T18:37:55ZengMDPI AGMolecules1420-30492021-02-01265127810.3390/molecules26051278Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel MyastheniaDenis Kudryavtsev0Anastasia Isaeva1Daria Barkova2Ekaterina Spirova3Renata Mukhutdinova4Igor Kasheverov5Victor Tsetlin6Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, RussiaShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, RussiaBiological Department, Lomonosov Moscow State University, 119991 Moscow, RussiaShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, RussiaShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, RussiaShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, RussiaShemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, RussiaSlow-channel congenital myasthenic syndromes (SCCMSs) are rare genetic diseases caused by mutations in muscle nicotinic acetylcholine receptor (nAChR) subunits. Most of the known SCCMS-associated mutations localize at the transmembrane region near the ion pore. Only two SCCMS point mutations are at the extracellular domains near the acetylcholine binding site, α1(G153S) being one of them. In this work, a combination of molecular dynamics, targeted mutagenesis, fluorescent Ca<sup>2+</sup> imaging and patch-clamp electrophysiology has been applied to G153S mutant muscle nAChR to investigate the role of hydrogen bonds formed by Ser 153 with C-loop residues near the acetylcholine-binding site. Introduction of L199T mutation to the C-loop in the vicinity of Ser 153 changed hydrogen bonds distribution, decreased acetylcholine potency (EC<sub>50</sub> 2607 vs. 146 nM) of the double mutant and decay kinetics of acetylcholine-evoked cytoplasmic Ca<sup>2+</sup> rise (τ 14.2 ± 0.3 vs. 34.0 ± 0.4 s). These results shed light on molecular mechanisms of nAChR activation-desensitization and on the involvement of such mechanisms in channelopathy genesis.https://www.mdpi.com/1420-3049/26/5/1278muscle nicotinic receptorgain-of-functionchannelopathyslow-channel congenital myastheniapatch-clampmolecular dynamics |
| spellingShingle | Denis Kudryavtsev Anastasia Isaeva Daria Barkova Ekaterina Spirova Renata Mukhutdinova Igor Kasheverov Victor Tsetlin Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia Molecules muscle nicotinic receptor gain-of-function channelopathy slow-channel congenital myasthenia patch-clamp molecular dynamics |
| title | Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia |
| title_full | Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia |
| title_fullStr | Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia |
| title_full_unstemmed | Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia |
| title_short | Point Mutations of Nicotinic Receptor α1 Subunit Reveal New Molecular Features of G153S Slow-Channel Myasthenia |
| title_sort | point mutations of nicotinic receptor α1 subunit reveal new molecular features of g153s slow channel myasthenia |
| topic | muscle nicotinic receptor gain-of-function channelopathy slow-channel congenital myasthenia patch-clamp molecular dynamics |
| url | https://www.mdpi.com/1420-3049/26/5/1278 |
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