Cryo-Electron microscopy for the study of self-assembled poly(ionic liquid) nanoparticles and protein supramolecular str
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INVITED ARTICLE
Cryo-Electron microscopy for the study of self-assembled poly(ionic liquid) nanoparticles and protein supramolecular structures Zdravko Kochovski 1
&
Guosong Chen 2
&
Jiayin Yuan 3 & Yan Lu 1,4
Received: 16 January 2020 / Revised: 8 April 2020 / Accepted: 9 April 2020 # The Author(s) 2020
Abstract Cryo-electron microscopy (cryo-EM) is a powerful structure determination technique that is well-suited to the study of protein and polymer self-assembly in solution. In contrast to conventional transmission electron microscopy (TEM) sample preparation, which often times involves drying and staining, the frozen-hydrated sample preparation allows the specimens to be kept and imaged in a state closest to their native one. Here, we give a short overview of the basic principles of Cryo-EM and review our results on applying it to the study of different protein and polymer self-assembled nanostructures. More specifically, we show how we have applied cryo-electron tomography (cryo-ET) to visualize the internal morphology of self-assembled poly(ionic liquid) nanoparticles and cryo-EM single particle analysis (SPA) to determine the three-dimensional (3D) structures of artificial protein microtubules. Keywords Self-assembly . Poly(ionic liquid) nanoparticles . Protein self-assembly . Cryo-electron microscopy . Single particle analysis . Cryo-electron tomography
Introduction
1
Institute of Electrochemical Energy Storage, Helmholtz-Zentrum Berlin für Materialien und Energie, 14109 Berlin, Germany
interest in recent years [3]. While TEM has established itself as the standard technique of choice for the structural and morphological analysis of self-assembled nanostructures, conventional TEM sample preparation often involves drying and staining steps which in many cases can affect the structure and morphology of the sample. In the case of cryo-EM, the aqueous samples are preserved in a frozen-hydrated state, closest to their native one, through vitrification and studied in the TEM at cryogenic temperatures. Cryo-EM is nowadays a powerful method for structure determination that has also been recently awarded with the 2017 Nobel Prize in Chemistry. The two main cryo-EM three-dimensional (3D) structure determination techniques are single-particle analysis (SPA) and cryo-electron tomography (cryo-ET). Here, we outline the basic principles of both techniques, including sample preparation, and give examples of how we have employed them in the study of protein and polymer self-assembled nanostructures.
2
The State Key Laboratory of Molecular Engineering of Polymers and Department of Macromolecular Science, Fudan University, Shanghai 200438, China
Sample preparation
3
Department of Materials and Environmental Chemistry, Stockholm University, SE-106 91 Stockholm, Sweden
4
Institute of Chemistry, University of Potsdam, 14476 Potsdam, Germany
Self-assembly of proteins plays a key role in many biological processes and has attracted substantial interest in recent years due to various potential applications in diagno
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