Haysom, SF, Machin, J, Whitehouse, JM et al. (9 more authors) (2023) Darobactin B Stabilises a Lateral‐Closed Conformation of the BAM Complex in E. coli Cells. Angewandte Chemie International Edition, 62 (34). ISSN 1433-7851
Abstract
The β-barrel assembly machinery (BAM complex) is essential for outer membrane protein (OMP) folding in Gram-negative bacteria, and represents a promising antimicrobial target. Several conformational states of BAM have been reported, but all have been obtained under conditions which lack the unique features and complexity of the outer membrane (OM). Here, we use Pulsed Electron-Electron Double Resonance (PELDOR, or DEER) spectroscopy distance measurements to interrogate the conformational ensemble of the BAM complex in E. coli cells. We show that BAM adopts a broad ensemble of conformations in the OM, while in the presence of the antibiotic darobactin B (DAR-B), BAM's conformational equilibrium shifts to a restricted ensemble consistent with the lateral closed state. Our in-cell PELDOR findings are supported by new cryoEM structures of BAM in the presence and absence of DAR-B. This work demonstrates the utility of PELDOR to map conformational changes in BAM within its native cellular environment.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
Keywords: | BAM; darobactin; In-cellEPR; PELDOR; cryoEM |
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) > Cryo EM, Image Processing (Leeds) The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology (Leeds) |
Funding Information: | Funder Grant number MRC (Medical Research Council) MR/P018491/1 BBSRC (Biotechnology & Biological Sciences Research Council) Not Known BBSRC (Biotechnology & Biological Sciences Research Council) BB/S018069/1 BBSRC (Biotechnology & Biological Sciences Research Council) BB/W019485/1 Royal Society RSRP\R1\211057 Wellcome Trust Not Known |
Depositing User: | Symplectic Publications |
Date Deposited: | 25 May 2023 11:36 |
Last Modified: | 25 Sep 2024 13:49 |
Published Version: | http://dx.doi.org/10.1002/anie.202218783 |
Status: | Published |
Publisher: | Wiley |
Identification Number: | 10.1002/anie.202218783 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:199208 |
Commentary/Response Threads
- Haysom, SF, Machin, J, Whitehouse, JM, Horne, JE, Fenn, K, Ma, Y, El Mkami, H, Böhringer, N, Schäberle, TF, Ranson, NA, Radford, SE and Pliotas, C Darobactin B Stabilises a Lateral‐Closed Conformation of the BAM Complex in E. coli Cells. (deposited 25 May 2023 11:36) [Currently Displayed]