Membrane Protein Cryo-EM: Cryo-Grid Optimization and Data Collection with Protein in Detergent

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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 languageEnglish
Title of host publicationExpression, Purification, and Structural Biology of Membrane Proteins
EditorsCamilo Perez, Timm Maier
Number of pages18
PublisherHumana Press
Publication date2020
ISBN (Print)978-1-0716-0372-7
ISBN (Electronic)978-1-0716-0373-4
Publication statusPublished - 2020
SeriesMethods in Molecular Biology

    Research areas

  • Cryo-electron microscopy, Data collection, Detergent, Grid preparation, Membrane protein, Negative staining, Structural biology, Vitrification

ID: 238002304