Membrane Protein Cryo-EM: Cryo-Grid Optimization and Data Collection with Protein in Detergent
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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.
Original language | English |
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Title of host publication | Expression, Purification, and Structural Biology of Membrane Proteins |
Editors | Camilo Perez, Timm Maier |
Number of pages | 18 |
Publisher | Humana Press |
Publication date | 2020 |
Pages | 227-244 |
ISBN (Print) | 978-1-0716-0372-7 |
ISBN (Electronic) | 978-1-0716-0373-4 |
DOIs | |
Publication status | Published - 2020 |
Series | Methods in Molecular Biology |
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Volume | 2127 |
ISSN | 1064-3745 |
- Cryo-electron microscopy, Data collection, Detergent, Grid preparation, Membrane protein, Negative staining, Structural biology, Vitrification
Research areas
ID: 238002304