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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