Preprint Review Version 1 Preserved in Portico This version is not peer-reviewed

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

Version 1 : Received: 16 August 2023 / Approved: 17 August 2023 / Online: 21 August 2023 (09:52:18 CEST)
Version 2 : Received: 21 September 2023 / Approved: 22 September 2023 / Online: 22 September 2023 (13:10:51 CEST)

A peer-reviewed article of this Preprint also exists.

Vénien-Bryan, C.; Fernandes, C.A.H. Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis. Int. J. Mol. Sci. 2023, 24, 14785. Vénien-Bryan, C.; Fernandes, C.A.H. Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis. Int. J. Mol. Sci. 2023, 24, 14785.

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 the last years, it was observed a huge increase in the number of MPs solved by cryo-EM SPA at a resolution better than 3.0 Å in the Protein Data Bank (PDB). However, sample preparation remains a significant challenge in the field. Here, we evaluated in the MPs solved by 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: 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.

Keywords

membrane proteins; cryo-electron microscopy; detergents; nanodiscs; amphipols; structural biology

Subject

Biology and Life Sciences, Biophysics

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