Parmar, M, Rawson, S orcid.org/0000-0002-2973-2630, Scarff, CA orcid.org/0000-0001-6168-0060 et al. (4 more authors) (2018) Using a SMALP platform to determine a sub-nm single particle cryo-EM membrane protein structure. BBA - Biomembranes, 1860 (2). pp. 378-383. ISSN 0005-2736
Abstract
The field of membrane protein structural biology has been revolutionized over the last few years with a number of high profile structures being solved using cryo-EM including Piezo, Ryanodine receptor, TRPV1 and the Glutamate receptor. Further developments in the EM field hold the promise of even greater progress in terms of greater resolution, which for membrane proteins is still typically within the 4–7 Å range. One advantage of a cryo-EM approach is the ability to study membrane proteins in more “native” like environments for example proteoliposomes, amphipols and nanodiscs. Recently, styrene maleic acid co-polymers (SMA) have been used to extract membrane proteins surrounded by native lipids (SMALPs) maintaining a more natural environment. We report here the structure of the Escherichia coli multidrug efflux transporter AcrB in a SMALP scaffold to sub-nm resolution, with the resulting map being consistent with high resolution crystal structures and other EM derived maps. However, both the C-terminal helix (TM12) and TM7 are poorly defined in the map. These helices are at the exterior of the helical bundle and form the greater interaction with the native lipids and SMA polymer and may represent a more dynamic region of the protein. This work shows the promise of using an SMA approach for single particle cryo-EM studies to provide sub-nm structures.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2017 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/BY/4.0/). |
Keywords: | SMALP; electron microscopy; membrane proteins; AcrB |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Animal Cell Biology (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cryo EM, Image Processing (Leeds) |
Funding Information: | Funder Grant number Wellcome Trust 108466/Z/15/Z |
Depositing User: | Symplectic Publications |
Date Deposited: | 09 Oct 2017 14:26 |
Last Modified: | 06 Oct 2018 00:39 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.bbamem.2017.10.005 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:122287 |
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