CW-EPR Spectroscopy and Site-Directed Spin Labeling to Study the Structural Dynamics of Ion Channels.
CW-EPR Spectroscopy and Site-Directed Spin Labeling to Study the Structural Dynamics of Ion Channels.
复制标题
CW-EPR 光谱和定点自旋标记研究离子通道的结构动力学。
DOI：
10.1007/978-1-4939-7362-0_21
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Cuello,LuisG
中科院分区：
文献类型：
--
作者：
Tilegenova,Cholpon;Elberson,BenjaminW;MarienCortes,D;Cuello,LuisG
连续波电子顺磁共振波谱（CW-EPR）和定点自旋标记（SDSL）是评估蛋白质结构动力学的有效实验方法(Hubbell et al.，生物化学学报，8(5):649-656,1998；中国生物医学工程学报（英文版），2016；科学通报，2009(5):344 - 344。这些技术特别有效地评估了嵌入脂质双分子层的整体膜蛋白的结构(Cortes et al., J Gen physil 117(2): 165-180, 2001；Cuello et al., Science 306(5695): 491-495, 2004；Dalmas等，结构18(7):868-878,2010；李等，中国科学院学报（自然科学版）112(44):E5926-5935, 2015；Perozo et al.，生物化学学报118(2):193 - 206,2001)，以及与生物功能相关的构象变化(Kazmier et al.，生物化学学报45:10 - 108,2016;Perozo et al.，生物科学285(5424):73 - 78,1999；[j] .中国生物医学工程学报，2016 (5):922-936；Dalmas et al.，《自然》（英文版）5:35 - 90,2014；[j] .中国科学：自然科学版，2008 (5):1023-1028,2005；Farrens et al., Science, 274(5288):768 - 770,1996；Perozo等，中华生物医学工程学报，5(6):459-469,1998；Perozo et al., Nature 418(6901): 942-948, 2002)。在本章中，我们描述了自旋标记、脂质体重构和典型细菌K+通道KcsA的CW-EPR测量的实用指南。
Continuous-wave electron paramagnetic resonance spectroscopy (CW-EPR) and site-directed spin labeling (SDSL) are proven experimental approaches to assess the structural dynamics of proteins in general (Hubbell et al., Curr Opin Struct Biol 8(5):649–656, 1998; Kazmier et al., Curr Opin Struct Biol 45:100–108, 2016; Perozo et al., Science 285(5424):73–78, 1999). These techniques have been particularly effective assessing the structure of integral membrane proteins embedded in a lipid bilayer (Cortes et al., J Gen Physiol 117(2):165–180, 2001; Cuello et al., Science 306(5695):491–495, 2004; Dalmas et al., Structure 18(7):868–878, 2010; Li et al., Proc Natl Acad Sci U S A 112(44):E5926–5935, 2015; Perozo et al., J Gen Physiol 118(2):193–206, 2001), as well as determining the conformational changes associated with their biological function (Kazmier et al., Curr Opin Struct Biol 45:100–108, 2016; Perozo et al., Science 285(5424):73–78, 1999; Arrigoni et al., Cell 164(5):922–936, 2016; Dalmas et al., Nat Commun 5:3590, 2014; Dong et al., Science 308(5724):1023–1028, 2005; Farrens et al., Science 274(5288):768–770, 1996; Perozo et al., Nat Struct Biol 5(6):459–469, 1998; Perozo et al., Nature 418(6901):942–948, 2002). In this chapter, we described a practical guide for the spin-labeling, liposome reconstitution, and CW-EPR measurements of the prototypical bacterial K+channel, KcsA.