PELDOR/DEER: An Electron Paramagnetic Resonance Method to Study Membrane Proteins in Lipid Bilayers.
PELDOR/DEER: An Electron Paramagnetic Resonance Method to Study Membrane Proteins in Lipid Bilayers.
复制标题
PELDOR/DEER：一种研究脂质双层膜蛋白的电子顺磁共振方法。
DOI：
10.1007/978-1-0716-0724-4_15
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Peter MF
中科院分区：
文献类型：
--
作者：
Peter MF
每个膜蛋白都与细胞膜的脂质环境密切相互作用。与多肽直接接触的环状脂质原则上可以被视为其结构的组成部分，类似于可溶性蛋白质的第一水合壳。因此，需要研究膜蛋白的结构，尤其是在尽可能接近其天然状态的条件下的构象灵活性。这可以通过将蛋白质重构为脂蛋白体、纳米圆盘或 bicelles 来实现。近年来，PELDOR/DEER光谱已被证明是研究此类人工膜环境中膜蛋白结构和功能的非常有用的方法。该技术补充了 X 射线晶体学和冷冻电镜，并且可以与几乎任何人造膜环境结合使用，在某些情况下甚至可以在天然膜中使用。在上述膜模拟物中，bicelles 目前最不常用于 PELDOR 研究，尽管它们具有一些优点，尤其是易于使用。在这里，我们提供了使用 PELDOR/DEER 光谱研究 bicelle 重组膜蛋白的分步方案。
Every membrane protein is involved in close interactions with the lipid environment of cellular membranes. The annular lipids, that are in direct contact with the polypeptide, can in principle be seen as an integral part of its structure, akin to the first hydration shell of soluble proteins. It is therefore desirable to investigate the structure of membrane proteins and especially their conformational flexibility under conditions that are as close as possible to their native state. This can be achieved by reconstituting the protein into proteoliposomes, nanodiscs, or bicelles. In recent years, PELDOR/DEER spectroscopy has proved to be a very useful method to study the structure and function of membrane proteins in such artificial membrane environments. The technique complements both X-ray crystallography and cryo-EM and can be used in combination with virtually any artificial membrane environment and under certain circumstances even in native membranes. Of the above-mentioned membrane mimics, bicelles are currently the least often used for PELDOR studies, although they offer some advantages, especially their ease of use. Here, we provide a step-by-step protocol for studying a bicelle reconstituted membrane protein with PELDOR/DEER spectroscopy.
登录
查看更多内容
影响因子：
15
作者：
Endeward, Burkhard;Butterwick, Joel A.;MacKinnon, Roderick;Prisner, Thomas F.
通讯作者：
Prisner, Thomas F.
影响因子：
3.4
作者：
Zou, Ping;Mchaourab, Hassane S.
通讯作者：
Mchaourab, Hassane S.
影响因子：
1
作者：
Hagelueken, Gregor;Ward, Richard;Naismith, James H.;Schiemann, Olav
通讯作者：
Schiemann, Olav
影响因子：
16.6
作者：
Joseph, Benesh;Sikora, Arthur;Bordignon, Enrica;Jeschke, Gunnar;Cafiso, David S.;Prisner, Thomas F.
通讯作者：
Prisner, Thomas F.
影响因子：
10.8
作者：
Orwick-Rydmark, Marcella;Lovett, Janet E.;Watts, Anthony
通讯作者：
Watts, Anthony