Electron microscopy as a critical tool in the determination of pore forming mechanisms in proteins.

Robert J C Gilbert

Methods in enzymology 2021

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Electron microscopy has consistently played an important role in the description of pore-forming protein systems. The discovery of pore-forming proteins has depended on visualization of the structural pores formed by their oligomeric protein complexes, and as electron microscopy has advanced technologically so has the degree of insight it has been able to give. This review considers a large number of published studies of pore-forming complexes in prepore and pore states determined using single-particle cryo-electron microscopy. Sample isolation and preparation, imaging and image analysis, structure determination and optimization of results are all discussed alongside challenges which pore-forming proteins particularly present. The review also considers the use made of cryo-electron tomography to study pores within their membrane environment and which will prove an increasingly important approach for the future. Copyright © 2021 Elsevier Inc. All rights reserved.

Citation

Robert J C Gilbert. Electron microscopy as a critical tool in the determination of pore forming mechanisms in proteins. Methods in enzymology. 2021;649:71-102