β1亚基的糖基化在毒素敏感性和BK通道的激活中起关键作用。

Xiaoli Wang, Qian Xiao, Yudan Zhu, Hong Qi, Dongxiao Qu, Yu Yao, Yuxiang Jia, Jingkan Guo, Jiwei Cheng, Yonghua Ji, Guoyi Li, Jie Tao
{"title":"β1亚基的糖基化在毒素敏感性和BK通道的激活中起关键作用。","authors":"Xiaoli Wang,&nbsp;Qian Xiao,&nbsp;Yudan Zhu,&nbsp;Hong Qi,&nbsp;Dongxiao Qu,&nbsp;Yu Yao,&nbsp;Yuxiang Jia,&nbsp;Jingkan Guo,&nbsp;Jiwei Cheng,&nbsp;Yonghua Ji,&nbsp;Guoyi Li,&nbsp;Jie Tao","doi":"10.1590/1678-9199-JVATITD-2020-0182","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The accessory β1 subunits, regulating the pharmacological and biophysical properties of BK channels, always undergo post-translational modifications, especially glycosylation. To date, it remains elusive whether the glycosylation contributes to the regulation of BK channels by β1 subunits.</p><p><strong>Methods: </strong>Herein, we combined the electrophysiological approach with molecular mutations and biochemical manipulation to investigate the function roles of N-glycosylation in β1 subunits.</p><p><strong>Results: </strong>The results show that deglycosylation of β1 subunits through double-site mutations (β1 N80A/N142A or β1 N80Q/N142Q) could significantly increase the inhibitory potency of iberiotoxin, a specific BK channel blocker. The deglycosylated channels also have a different sensitivity to martentoxin, another BK channel modulator with some remarkable effects as reported before. On the contrary to enhancing effects of martentoxin on glycosylated BK channels under the presence of cytoplasmic Ca<sup>2+</sup>, deglycosylated channels were not affected by the toxin. However, the deglycosylated channels were surprisingly inhibited by martentoxin under the absence of cytoplasmic Ca<sup>2+</sup>, while the glycosylated channels were not inhibited under this same condition. In addition, wild type BK (α+β1) channels treated with PNGase F also showed the same trend of pharmacological results to the mutants. Similar to this modulation of glycosylation on BK channel pharmacology, the deglycosylated forms of the channels were activated at a faster speed than the glycosylated ones. However, the V<sub>1/2</sub> and slope were not changed by the glycosylation.</p><p><strong>Conclusion: </strong>The present study reveals that glycosylation is an indispensable determinant of the modulation of β1-subunit on BK channel pharmacology and its activation. The loss of glycosylation of β1 subunits could lead to the dysfunction of BK channel, resulting in a pathological state.</p>","PeriodicalId":520810,"journal":{"name":"The journal of venomous animals and toxins including tropical diseases","volume":" ","pages":"e20200182"},"PeriodicalIF":0.0000,"publicationDate":"2021-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8183112/pdf/","citationCount":"3","resultStr":"{\"title\":\"Glycosylation of β1 subunit plays a pivotal role in the toxin sensitivity and activation of BK channels.\",\"authors\":\"Xiaoli Wang,&nbsp;Qian Xiao,&nbsp;Yudan Zhu,&nbsp;Hong Qi,&nbsp;Dongxiao Qu,&nbsp;Yu Yao,&nbsp;Yuxiang Jia,&nbsp;Jingkan Guo,&nbsp;Jiwei Cheng,&nbsp;Yonghua Ji,&nbsp;Guoyi Li,&nbsp;Jie Tao\",\"doi\":\"10.1590/1678-9199-JVATITD-2020-0182\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The accessory β1 subunits, regulating the pharmacological and biophysical properties of BK channels, always undergo post-translational modifications, especially glycosylation. To date, it remains elusive whether the glycosylation contributes to the regulation of BK channels by β1 subunits.</p><p><strong>Methods: </strong>Herein, we combined the electrophysiological approach with molecular mutations and biochemical manipulation to investigate the function roles of N-glycosylation in β1 subunits.</p><p><strong>Results: </strong>The results show that deglycosylation of β1 subunits through double-site mutations (β1 N80A/N142A or β1 N80Q/N142Q) could significantly increase the inhibitory potency of iberiotoxin, a specific BK channel blocker. The deglycosylated channels also have a different sensitivity to martentoxin, another BK channel modulator with some remarkable effects as reported before. On the contrary to enhancing effects of martentoxin on glycosylated BK channels under the presence of cytoplasmic Ca<sup>2+</sup>, deglycosylated channels were not affected by the toxin. However, the deglycosylated channels were surprisingly inhibited by martentoxin under the absence of cytoplasmic Ca<sup>2+</sup>, while the glycosylated channels were not inhibited under this same condition. In addition, wild type BK (α+β1) channels treated with PNGase F also showed the same trend of pharmacological results to the mutants. Similar to this modulation of glycosylation on BK channel pharmacology, the deglycosylated forms of the channels were activated at a faster speed than the glycosylated ones. However, the V<sub>1/2</sub> and slope were not changed by the glycosylation.</p><p><strong>Conclusion: </strong>The present study reveals that glycosylation is an indispensable determinant of the modulation of β1-subunit on BK channel pharmacology and its activation. The loss of glycosylation of β1 subunits could lead to the dysfunction of BK channel, resulting in a pathological state.</p>\",\"PeriodicalId\":520810,\"journal\":{\"name\":\"The journal of venomous animals and toxins including tropical diseases\",\"volume\":\" \",\"pages\":\"e20200182\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2021-06-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8183112/pdf/\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The journal of venomous animals and toxins including tropical diseases\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1590/1678-9199-JVATITD-2020-0182\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The journal of venomous animals and toxins including tropical diseases","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1590/1678-9199-JVATITD-2020-0182","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3

摘要

背景:辅助β1亚基调节BK通道的药理学和生物物理性质,经常发生翻译后修饰,特别是糖基化。迄今为止,糖基化是否参与β1亚基对BK通道的调节尚不清楚。方法:采用电生理方法、分子突变和生化操作相结合的方法,研究β1亚基n -糖基化的功能作用。结果:结果表明,通过双位点突变(β1 N80A/N142A或β1 N80Q/N142Q)使β1亚基去糖基化可显著提高特异性BK通道阻滞剂iberiotoxin的抑制效能。去糖基化通道对马氏毒素也有不同的敏感性,马氏毒素是另一种BK通道调节剂,具有一些显着的作用,如前所述。相反,在细胞质Ca2+存在下,马氏毒素对糖基化BK通道的作用增强,去糖基化通道不受毒素的影响。然而,在缺乏细胞质Ca2+的情况下,去糖基化通道令人惊讶地受到马氏毒素的抑制,而糖基化通道在相同条件下没有受到抑制。此外,PNGase F处理野生型BK (α+β1)通道对突变体也表现出相同的药理结果趋势。与糖基化对BK通道药理学的调节类似,去糖基化形式的通道比糖基化形式的通道以更快的速度激活。然而,V1/2和斜率不受糖基化的影响。结论:糖基化是β1亚基调控BK通道药理学及其激活的重要决定因素。β1亚基糖基化缺失可导致BK通道功能障碍,导致病理状态。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Glycosylation of β1 subunit plays a pivotal role in the toxin sensitivity and activation of BK channels.

Glycosylation of β1 subunit plays a pivotal role in the toxin sensitivity and activation of BK channels.

Glycosylation of β1 subunit plays a pivotal role in the toxin sensitivity and activation of BK channels.

Glycosylation of β1 subunit plays a pivotal role in the toxin sensitivity and activation of BK channels.

Background: The accessory β1 subunits, regulating the pharmacological and biophysical properties of BK channels, always undergo post-translational modifications, especially glycosylation. To date, it remains elusive whether the glycosylation contributes to the regulation of BK channels by β1 subunits.

Methods: Herein, we combined the electrophysiological approach with molecular mutations and biochemical manipulation to investigate the function roles of N-glycosylation in β1 subunits.

Results: The results show that deglycosylation of β1 subunits through double-site mutations (β1 N80A/N142A or β1 N80Q/N142Q) could significantly increase the inhibitory potency of iberiotoxin, a specific BK channel blocker. The deglycosylated channels also have a different sensitivity to martentoxin, another BK channel modulator with some remarkable effects as reported before. On the contrary to enhancing effects of martentoxin on glycosylated BK channels under the presence of cytoplasmic Ca2+, deglycosylated channels were not affected by the toxin. However, the deglycosylated channels were surprisingly inhibited by martentoxin under the absence of cytoplasmic Ca2+, while the glycosylated channels were not inhibited under this same condition. In addition, wild type BK (α+β1) channels treated with PNGase F also showed the same trend of pharmacological results to the mutants. Similar to this modulation of glycosylation on BK channel pharmacology, the deglycosylated forms of the channels were activated at a faster speed than the glycosylated ones. However, the V1/2 and slope were not changed by the glycosylation.

Conclusion: The present study reveals that glycosylation is an indispensable determinant of the modulation of β1-subunit on BK channel pharmacology and its activation. The loss of glycosylation of β1 subunits could lead to the dysfunction of BK channel, resulting in a pathological state.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信