Collinson Ian, Vonck Janet, Hizlan Dilem
School of Biochemistry, University of Bristol, Bristol, UK.
Methods Mol Biol. 2013;1033:47-65. doi: 10.1007/978-1-62703-487-6_4.
Electron crystallography is a powerful technique for studying the structure and function of membrane proteins, not only in the ground state, but also in active conformations. When combined with high-resolution structures obtained by X-ray crystallography, electron crystallography can provide insights into the mechanism of the protein. In this chapter we discuss obtaining a three-dimensional map of membrane proteins by electron crystallography and how to combine these maps with atomic resolution models in order to study the function of membrane proteins. We argue that this approach is particularly powerful as it combines the high resolution attainable by X-ray crystallography with the visualization of the subject in the near-native environment of the membrane, by electron cryo-microscopy. This point has been illustrated by the analysis of the protein translocation complex SecYEG.
电子晶体学是研究膜蛋白结构和功能的一种强大技术，不仅可用于研究其基态，还可用于研究其活性构象。当与通过X射线晶体学获得的高分辨率结构相结合时，电子晶体学能够深入了解蛋白质的作用机制。在本章中，我们将讨论如何通过电子晶体学获得膜蛋白的三维图谱，以及如何将这些图谱与原子分辨率模型相结合，以研究膜蛋白的功能。我们认为这种方法特别有效，因为它将X射线晶体学可达到的高分辨率与通过电子冷冻显微镜在膜的近天然环境中对研究对象的可视化相结合。对蛋白质转运复合物SecYEG的分析就说明了这一点。
