Sample Preparation for Electron Cryo-Microscopy of Macromolecular Machines - Archive ouverte HAL
Chapitre D'ouvrage Année : 2024

Sample Preparation for Electron Cryo-Microscopy of Macromolecular Machines

Résumé

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.
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Dates et versions

hal-04608691 , version 1 (11-06-2024)

Identifiants

Citer

Aurélien Deniaud, Burak V. Kabasakal, Joshua C. Bufton, Christiane Schaffitzel. Sample Preparation for Electron Cryo-Microscopy of Macromolecular Machines. M. Cristina Vega, Francisco J. Fernández. Advanced Technologies for Protein Complex Production and Characterization vol.2; Part of the book series: Advances in Experimental Medicine and Biology (AEMB, volume 1453), 1453, SPRINGER INTERNATIONAL PUBLISHING AGGEWERBESTRASSE 11, CHAM CH-6330, SWITZERLAND, pp.173-190, 2024, ⟨10.1007/978-3-031-52193-5_12⟩. ⟨hal-04608691⟩
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