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- Title
Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis.
- Authors
Higgins, Anna J.; Flynn, Alex J.; Marconnet, Anaïs; Musgrove, Laura J.; Postis, Vincent L. G.; Lippiat, Jonathan D.; Chung, Chun-wa; Ceska, Tom; Zoonens, Manuela; Sobott, Frank; Muench, Stephen P.
- Abstract
Membrane proteins are essential for cellular growth, signalling and homeostasis, making up a large proportion of therapeutic targets. However, the necessity for a solubilising agent to extract them from the membrane creates challenges in their structural and functional study. Although amphipols have been very effective for single-particle electron cryo-microscopy (cryoEM) and mass spectrometry, they rely on initial detergent extraction before exchange into the amphipol environment. Therefore, circumventing this pre-requirement would be a big advantage. Here we use an alternative type of amphipol: a cycloalkane-modified amphiphile polymer (CyclAPol) to extract Escherichia coli AcrB directly from the membrane and demonstrate that the protein can be isolated in a one-step purification with the resultant cryoEM structure achieving 3.2 Å resolution. Together this work shows that cycloalkane amphipols provide a powerful approach for the study of membrane proteins, allowing native extraction and high-resolution structure determination by cryoEM. Higgins et al. present a cycloalkane-modified amphiphilic polymer that can provide direct extraction of membrane proteins for Cryo-EM analysis. They show its utility by extracting and solving the structure of AcrB to a high resolution of 3.2 Å by single particle cryo-EM.
- Subjects
MEMBRANE proteins; PROTEIN analysis; POLYMERS; DRUG target; CELL growth
- Publication
Communications Biology, 2021, Vol 4, Issue 1, p1
- ISSN
2399-3642
- Publication type
Article
- DOI
10.1038/s42003-021-02834-3