Journal
COMMUNICATIONS BIOLOGY
Volume 4, Issue 1, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/s42003-021-02834-3
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Funding
- University of Leeds (UoL ABSL award)
- Wellcome Trust [108466/Z/15/Z]
- BBSRC [BB/R018561/1]
- Centre National de la Recherche Scientifique (CNRS)
- Universite de Paris (Universite Paris 7)
- 'Initiative d'Excellence' programme from the French State (Grant 'DYNAMO') [ANR-11-LABX-0011-01]
- BBSRC [BB/R018561/1] Funding Source: UKRI
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The study presents a novel approach using cycloalkane-modified amphiphilic polymer for direct extraction of membrane proteins, showcasing its utility in solving the structure of AcrB to a high resolution of 3.2 angstrom by single particle cryo-EM.
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 angstrom 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 angstrom by single particle cryo-EM.
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