School of Molecular Sciences, Arizona State University, Tempe, AZ, USA.
The Biodesign Institute, Arizona State University, Tempe, AZ, USA.
Methods Mol Biol. 2021;2215:247-265. doi: 10.1007/978-1-0716-0966-8_11.
Electron crystallography is a unique tool to study membrane protein structures and lipid-protein interactions in their native-like environments. Two-dimensional (2D) protein crystallization enables the lipids immobilized by the proteins, and the generated high-resolution density map allows us to model the atomic coordinates of the surrounding lipids to study lipid-protein interaction. This protocol describes the sample preparation for electron crystallographic studies, including back-injection method and carbon sandwich method. The protocols of data collection for electron crystallography, including electron imaging and diffraction, of the 2D membrane crystal will be followed.
电子晶体学是一种独特的工具,可用于研究天然环境中膜蛋白结构和脂质-蛋白相互作用。二维(2D)蛋白质结晶能够使蛋白质固定化的脂质固定下来,生成的高分辨率密度图使我们能够模拟周围脂质的原子坐标,从而研究脂质-蛋白相互作用。本方案介绍了电子晶体学研究的样品制备,包括背注入法和碳夹层法。随后将介绍 2D 膜晶体的电子晶体学数据收集方案,包括电子成像和衍射。