Membrane Protein Cryo-EM: Cryo-Grid Optimization and Data Collection with Protein in Detergent.
Methods Mol Biol
; 2127: 227-244, 2020.
Article
em En
| MEDLINE
| ID: mdl-32112326
ABSTRACT
Cryo-electron microscopy (cryo-EM) is a powerful tool for investigating the structure of macromolecules under near-native conditions. Especially in the context of membrane proteins, this technique has allowed researchers to obtain structural information at a previously unattainable level of detail. Specimen preparation remains the bottleneck of most cryo-EM research projects, with membrane proteins representing particularly challenging targets of investigation due to their universal requirement for detergents or other solubilizing agents. Here we describe preparation of negative staining and cryo-EM grids and downstream data collection of membrane proteins in detergent, by far the most common solubilization agent. This protocol outlines a quick and straightforward procedure for screening and determining the structure of a membrane protein of interest under biologically relevant conditions.
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Base de dados:
MEDLINE
Assunto principal:
Coleta de Dados
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Microscopia Crioeletrônica
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Detergentes
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Proteínas de Membrana
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article