Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor
Recently, the muscle-type nicotinic acetylcholine receptors (nAChRs) have been pursued as a potential target of several diseases, including myogenic disorders, muscle dystrophies and myasthenia gravis, etc. α-conotoxin GI isolated from <i>Conus geographus</i> selectively and pot...
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doaj-e523b49edef74d5f8c6f77c7dda777fe2020-11-24T20:44:54ZengMDPI AGMarine Drugs1660-33972018-12-01161250710.3390/md16120507md16120507Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine ReceptorJiong Ning0Rui Li1Jie Ren2Dongting Zhangsun3Xiaopeng Zhu4Yong Wu5Sulan Luo6Key Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaKey Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaKey Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaKey Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaKey Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaKey Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaKey Laboratory of Tropical Biological Resources, Ministry of Education, Key Lab for Marine Drugs of Haikou, Hainan University, Haikou 570228, Hainan, ChinaRecently, the muscle-type nicotinic acetylcholine receptors (nAChRs) have been pursued as a potential target of several diseases, including myogenic disorders, muscle dystrophies and myasthenia gravis, etc. α-conotoxin GI isolated from <i>Conus geographus</i> selectively and potently inhibited the muscle-type nAChRs which can be developed as a tool to study them. Herein, alanine scanning mutagenesis was used to reveal the structure⁻activity relationship (SAR) between GI and mouse α1β1δε nAChRs. The Pro<sup>5</sup>, Gly<sup>8</sup>, Arg<sup>9</sup>, and Tyr<sup>11</sup> were proved to be the critical residues for receptor inhibiting as the alanine (Ala) replacement led to a significant potency loss on mouse α1β1δε nAChR. On the contrary, substituting Asn<sup>4</sup>, His<sup>10</sup> and Ser<sup>12</sup> with Ala respectively did not affect its activity. Interestingly, the [E1A] GI analogue exhibited a three-fold potency for mouse α1β1δε nAChR, whereas it obviously decreased potency at rat α9α10 nAChR compared to wildtype GI. Molecular dynamic simulations also suggest that loop2 of GI significantly affects the interaction with α1β1δε nAChR, and Tyr<sup>11</sup> of GI is a critical residue binding with three hydrophobic amino acids of the δ subunit, including Leu<sup>93</sup>, Tyr<sup>95</sup> and Leu<sup>103</sup>. Our research elucidates the interaction of GI and mouse α1β1δε nAChR in detail that will help to develop the novel analogues of GI.https://www.mdpi.com/1660-3397/16/12/507muscle-type nAChRsα-conotoxin GIstructure-activity relationshipelectrophysiological studymolecular dynamic simulations |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jiong Ning Rui Li Jie Ren Dongting Zhangsun Xiaopeng Zhu Yong Wu Sulan Luo |
spellingShingle |
Jiong Ning Rui Li Jie Ren Dongting Zhangsun Xiaopeng Zhu Yong Wu Sulan Luo Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor Marine Drugs muscle-type nAChRs α-conotoxin GI structure-activity relationship electrophysiological study molecular dynamic simulations |
author_facet |
Jiong Ning Rui Li Jie Ren Dongting Zhangsun Xiaopeng Zhu Yong Wu Sulan Luo |
author_sort |
Jiong Ning |
title |
Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor |
title_short |
Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor |
title_full |
Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor |
title_fullStr |
Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor |
title_full_unstemmed |
Alanine-Scanning Mutagenesis of α-Conotoxin GI Reveals the Residues Crucial for Activity at the Muscle Acetylcholine Receptor |
title_sort |
alanine-scanning mutagenesis of α-conotoxin gi reveals the residues crucial for activity at the muscle acetylcholine receptor |
publisher |
MDPI AG |
series |
Marine Drugs |
issn |
1660-3397 |
publishDate |
2018-12-01 |
description |
Recently, the muscle-type nicotinic acetylcholine receptors (nAChRs) have been pursued as a potential target of several diseases, including myogenic disorders, muscle dystrophies and myasthenia gravis, etc. α-conotoxin GI isolated from <i>Conus geographus</i> selectively and potently inhibited the muscle-type nAChRs which can be developed as a tool to study them. Herein, alanine scanning mutagenesis was used to reveal the structure⁻activity relationship (SAR) between GI and mouse α1β1δε nAChRs. The Pro<sup>5</sup>, Gly<sup>8</sup>, Arg<sup>9</sup>, and Tyr<sup>11</sup> were proved to be the critical residues for receptor inhibiting as the alanine (Ala) replacement led to a significant potency loss on mouse α1β1δε nAChR. On the contrary, substituting Asn<sup>4</sup>, His<sup>10</sup> and Ser<sup>12</sup> with Ala respectively did not affect its activity. Interestingly, the [E1A] GI analogue exhibited a three-fold potency for mouse α1β1δε nAChR, whereas it obviously decreased potency at rat α9α10 nAChR compared to wildtype GI. Molecular dynamic simulations also suggest that loop2 of GI significantly affects the interaction with α1β1δε nAChR, and Tyr<sup>11</sup> of GI is a critical residue binding with three hydrophobic amino acids of the δ subunit, including Leu<sup>93</sup>, Tyr<sup>95</sup> and Leu<sup>103</sup>. Our research elucidates the interaction of GI and mouse α1β1δε nAChR in detail that will help to develop the novel analogues of GI. |
topic |
muscle-type nAChRs α-conotoxin GI structure-activity relationship electrophysiological study molecular dynamic simulations |
url |
https://www.mdpi.com/1660-3397/16/12/507 |
work_keys_str_mv |
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