利用冷冻电镜（Cryo-EM）方法的微束电子衍射（MicroED）进行蛋白质和小分子结构测定。
Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED.
机构信息
Department of Biological Chemistry, University of California Los Angeles, Los Angeles, CA, USA.
Department of Physiology, University of California Los Angeles, Los Angeles, CA, USA.
出版信息
Methods Mol Biol. 2021;2305:323-342. doi: 10.1007/978-1-0716-1406-8_16.
Microcrystal Electron Diffraction (MicroED) is the newest cryo-electron microscopy (cryo-EM) method, with over 70 protein, peptide, and several small organic molecule structures already determined. In MicroED, micro- or nanocrystalline samples in solution are deposited on electron microscopy grids and examined in a cryo-electron microscope, ideally under cryogenic conditions. Continuous rotation diffraction data are collected and then processed using conventional X-ray crystallography programs. The protocol outlined here details how to obtain and identify the nanocrystals, how to set up the microscope for screening and for MicroED data collection, and how to collect and process data to complete high-resolution structures. For well-behaving crystals with high-resolution diffraction in cryo-EM, the entire process can be achieved in less than an hour.
微晶体电子衍射（MicroED）是最新的冷冻电镜（cryo-EM）方法，已经有超过 70 种蛋白质、肽和几种小分子有机分子结构被确定。在 MicroED 中，溶液中的微晶体或纳米晶体样品沉积在电子显微镜载网上，并在冷冻电子显微镜下进行检查，理想情况下在低温条件下进行。连续旋转衍射数据被收集，然后使用传统的 X 射线晶体学程序进行处理。这里概述的方案详细说明了如何获得和识别纳米晶体，如何设置显微镜进行筛选和 MicroED 数据收集，以及如何收集和处理数据以完成高分辨率结构。对于在 cryo-EM 中具有高分辨率衍射的良好晶体，整个过程可以在不到一个小时内完成。
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