Sample Preparation and Data Collection for Electron Crystallographic Studies on Membrane Protein Structures and Lipid–Protein Interaction

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.