@misc{da6cc20df433495fa1f135aa5eb64a99,
title = "Cycloalkane-modified amphiphilic polymers provide direct extraction of membrane proteins for CryoEM analysis",
abstract = "Membrane proteins are essential for cellular growth and homeostasis, making up a large proportion of therapeutic targets. However, the necessity for a solubilising agent to extract them from the membrane creates significant 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 significant advantage. Here we use a novel 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 {\AA} resolution. Together this work shows that cycloalkane amphipols provide a powerful detergent-free approach for the study of membrane proteins allowing native extraction and high-resolution structure determination by cryoEM.",
keywords = "CryoEM, membrane protein, amphipol, detergent",
author = "Anna Higgins and Alex Flynn and Ana{\"i}s Marconnet and Laura Musgrove and Vincent Postis and Jonathan Lippiat and Chun-Wa Chung and Tom Ceska and Manuela Zoonens and Frank Sobott and Stephen Muench",
note = "The FEI Titan Krios microscopes were funded by the University of Leeds (UoL ABSL award) and Wellcome Trust (108466/Z/15/Z). This work and AJH was funded through a BBSRC grant (BB/R018561/1/). The amphipol development in IBPC Paris was supported by the Centre National de la Recherche Scientifique (CNRS), Universit{\'e} de Paris (Universit{\'e} Paris 7), and the “Initiative d{\textquoteright}Excellence” program from the French State (Grant “DYNAMO”, ANR-11-LABX-0011-01)",
year = "2021",
month = jan,
day = "15",
doi = "10.21203/rs.3.rs-131488/v1",
language = "English",
publisher = "Research Square",
address = "United States",
type = "Other",
}