Sample Preparation for Electron Cryo-Microscopy of Macromolecular Machines

Adv Exp Med Biol. 2024;3234:173-190. doi: 10.1007/978-3-031-52193-5_12.ABSTRACTHigh-resolution structure determination by electron cryo-microscopy underwent a step change in recent years. This now allows study of challenging samples which previously were inaccessible for structure determination, including membrane proteins. These developments shift the focus in the field to the next bottlenecks which are high-quality sample preparations. While the amounts of sample required for cryo-EM are relatively small, sample quality is the key challenge. Sample quality is influenced by the stability of complexes which depends on buffer composition, inherent flexibility of the sample, and the method of solubilization from the membrane for membrane proteins. It further depends on the choice of sample support, grid pre-treatment and cryo-grid freezing protocol. Here, we discuss various widely applicable approaches to improve sample quality for structural analysis by cryo-EM.PMID:38507207 | DOI:10.1007/978-3-031-52193-5_12

Source: Advances in Experimental Medicine and Biology - March 20, 2024 Category: Research Authors: Aur élien Deniaud Burak V Kabasakal Joshua C Bufton Christiane Schaffitzel Source Type: research