Gonen Tamir
Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
Methods Mol Biol. 2013;955:153-69. doi: 10.1007/978-1-62703-176-9_9.
A number of atomic-resolution structures of membrane proteins (better than 3Å resolution) have been determined recently by electron crystallography. While this technique was established more than 40 years ago, it is still in its infancy with regard to the two-dimensional (2D) crystallization, data collection, data analysis, and protein structure determination. In terms of data collection, electron crystallography encompasses both image acquisition and electron diffraction data collection. Other chapters in this volume outline protocols for image collection and analysis. This chapter, however, outlines detailed protocols for data collection by electron diffraction. These include microscope setup, electron diffraction data collection, and troubleshooting.
近年来,通过电子晶体学已经确定了许多膜蛋白的原子分辨率结构(分辨率优于3Å)。尽管这项技术在40多年前就已建立,但在二维(2D)结晶、数据收集、数据分析和蛋白质结构测定方面仍处于起步阶段。在数据收集方面,电子晶体学包括图像采集和电子衍射数据收集。本卷中的其他章节概述了图像采集和分析的方案。然而,本章概述了通过电子衍射进行数据收集的详细方案。这些包括显微镜设置、电子衍射数据收集和故障排除。