稳定功能性膜蛋白的洗涤剂的筛选

Q1 Biochemistry, Genetics and Molecular Biology Current Protocols in Protein Science Pub Date : 2018-07-18 DOI:10.1002/cpps.59
Guillaume Lenoir, Thibaud Dieudonné, Anaïs Lamy, Maylis Lejeune, José-Luis Vazquez-Ibar, Cédric Montigny
{"title":"稳定功能性膜蛋白的洗涤剂的筛选","authors":"Guillaume Lenoir,&nbsp;Thibaud Dieudonné,&nbsp;Anaïs Lamy,&nbsp;Maylis Lejeune,&nbsp;José-Luis Vazquez-Ibar,&nbsp;Cédric Montigny","doi":"10.1002/cpps.59","DOIUrl":null,"url":null,"abstract":"<p>Membrane protein studies usually require use of detergents to extract and isolate proteins from membranes and manipulate them in a soluble context for their functional or structural characterization. However, solubilization with detergent may interfere with MP stability and may directly affect MP function or structure. Moreover, detergent properties can be affected such as critical micellar concentration (CMC) can be affected by the experimental conditions. Consequently, the experimenter must pay attention to both the protein and the behavior of the detergent. This article provides a convenient protocol for estimating the CMC of detergents in given experimental conditions. Then, it presents two protocols aimed at monitoring the function of a membrane protein in the presence of detergent. Such experiments may help to test various detergents for their inactivating or stabilizing effects on long incubation times, ranging from few hours to some days. © 2018 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":10866,"journal":{"name":"Current Protocols in Protein Science","volume":"93 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpps.59","citationCount":"7","resultStr":"{\"title\":\"Screening of Detergents for Stabilization of Functional Membrane Proteins\",\"authors\":\"Guillaume Lenoir,&nbsp;Thibaud Dieudonné,&nbsp;Anaïs Lamy,&nbsp;Maylis Lejeune,&nbsp;José-Luis Vazquez-Ibar,&nbsp;Cédric Montigny\",\"doi\":\"10.1002/cpps.59\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Membrane protein studies usually require use of detergents to extract and isolate proteins from membranes and manipulate them in a soluble context for their functional or structural characterization. However, solubilization with detergent may interfere with MP stability and may directly affect MP function or structure. Moreover, detergent properties can be affected such as critical micellar concentration (CMC) can be affected by the experimental conditions. Consequently, the experimenter must pay attention to both the protein and the behavior of the detergent. This article provides a convenient protocol for estimating the CMC of detergents in given experimental conditions. Then, it presents two protocols aimed at monitoring the function of a membrane protein in the presence of detergent. Such experiments may help to test various detergents for their inactivating or stabilizing effects on long incubation times, ranging from few hours to some days. © 2018 by John Wiley &amp; Sons, Inc.</p>\",\"PeriodicalId\":10866,\"journal\":{\"name\":\"Current Protocols in Protein Science\",\"volume\":\"93 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2018-07-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cpps.59\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Protocols in Protein Science\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpps.59\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Protein Science","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpps.59","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 7

摘要

膜蛋白研究通常需要使用洗涤剂从膜中提取和分离蛋白质,并在可溶性环境中对其进行功能或结构表征。然而,与洗涤剂的增溶可能会干扰MP的稳定性,并可能直接影响MP的功能或结构。此外,实验条件会影响洗涤剂的性能,如临界胶束浓度(CMC)。因此,实验人员必须同时注意蛋白质和洗涤剂的行为。本文提供了一种在给定实验条件下估计洗涤剂CMC的简便方法。然后,它提出了两种方案,旨在监测膜蛋白的功能在洗涤剂的存在。这样的实验可以帮助测试各种洗涤剂在长孵育时间(从几小时到几天)内的灭活或稳定效果。©2018 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Screening of Detergents for Stabilization of Functional Membrane Proteins

Membrane protein studies usually require use of detergents to extract and isolate proteins from membranes and manipulate them in a soluble context for their functional or structural characterization. However, solubilization with detergent may interfere with MP stability and may directly affect MP function or structure. Moreover, detergent properties can be affected such as critical micellar concentration (CMC) can be affected by the experimental conditions. Consequently, the experimenter must pay attention to both the protein and the behavior of the detergent. This article provides a convenient protocol for estimating the CMC of detergents in given experimental conditions. Then, it presents two protocols aimed at monitoring the function of a membrane protein in the presence of detergent. Such experiments may help to test various detergents for their inactivating or stabilizing effects on long incubation times, ranging from few hours to some days. © 2018 by John Wiley & Sons, Inc.

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
Current Protocols in Protein Science
Current Protocols in Protein Science Biochemistry, Genetics and Molecular Biology-Biochemistry
自引率
0.00%
发文量
0
期刊介绍: With the mapping of the human genome, more and more researchers are exploring protein structures and functions in living organisms. Current Protocols in Protein Science provides protein scientists, biochemists, molecular biologists, geneticists, and others with the first comprehensive suite of protocols for this growing field.
期刊最新文献
Issue Information De Novo Protein Design Using the Blueprint Builder in Rosetta Methods for Expression of Recombinant Proteins Using a Pichia pastoris Cell-Free System Histone Purification Combined with High-Resolution Mass Spectrometry to Examine Histone Post-Translational Modifications and Histone Variants in Caenorhabditis elegans Issue Information
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1