Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis - Sorbonne Université Access content directly
Journal Articles (Review Article) International Journal of Molecular Sciences Year : 2023

Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis

Abstract

Single-particle cryo-electron microscopy (cryo-EM SPA) has recently emerged as an exceptionally well-suited technique for determining the structure of membrane proteins (MPs). Indeed, in recent years, huge increase in the number of MPs solved via cryo-EM SPA at a resolution better than 3.0 Å in the Protein Data Bank (PDB) has been observed. However, sample preparation remains a significant challenge in the field. Here, we evaluated the MPs solved using cryo-EM SPA deposited in the PDB in the last two years at a resolution below 3.0 Å. The most critical parameters for sample preparation are as follows: (i) the surfactant used for protein extraction from the membrane, (ii) the surfactant, amphiphiles, nanodiscs or other molecules present in the vitrification step, (iii) the vitrification method employed, and (iv) the type of grids used. The aim is not to provide a definitive answer on the optimal sample conditions for cryo-EM SPA of MPs but rather assess the current trends in the MP structural biology community towards obtaining high-resolution cryo-EM structures.
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Dates and versions

hal-04229254 , version 1 (05-10-2023)

Identifiers

  • HAL Id : hal-04229254 , version 1

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Catherine Vénien-Bryan, Carlos A.H. Fernandes. Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis. International Journal of Molecular Sciences, 2023, 24. ⟨hal-04229254⟩
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