Sample Preparation for Electron Cryo-Microscopy of Macromolecular Machines

Date

2024-01-01

Author

Deniaud, Aurelien
Kabasakal, Burak Veli
Bufton, Joshua C.
Schaffitzel, Christiane

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High-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.

URI

https://hdl.handle.net/11511/116751

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Department of Biology, Article