Journal article

Authors list: Haysom, Samuel F.; Machin, Jonathan; Whitehouse, James M.; Horne, Jim E.; Fenn, Katherine; Ma, Yue; El Mkami, Hassane; Böhringer, Nils; Schäberle, Till F.; Ranson, Neil A.; Radford, Sheena E.; Pliotas, Christos

Publication year: 2023

Journal: Angewandte Chemie International Edition

Volume number: 62

Issue number: 34

ISSN: 1433-7851

eISSN: 1521-3773

Open access status: Hybrid

DOI Link: https://doi.org/10.1002/anie.202218783

Publisher: Wiley

Abstract: 
The beta-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.

Harvard Citation style: Haysom, S., Machin, J., Whitehouse, J., Horne, J., Fenn, K., Ma, Y., et al. (2023) Darobactin B Stabilises a Lateral-Closed Conformation of the BAM Complex in E. coli Cells, Angewandte Chemie International Edition, 62(34), Article e202218783. https://doi.org/10.1002/anie.202218783

APA Citation style: Haysom, S., Machin, J., Whitehouse, J., Horne, J., Fenn, K., Ma, Y., El Mkami, H., Böhringer, N., Schäberle, T., Ranson, N., Radford, S., & Pliotas, C. (2023). Darobactin B Stabilises a Lateral-Closed Conformation of the BAM Complex in E. coli Cells. Angewandte Chemie International Edition. 62(34), Article e202218783. https://doi.org/10.1002/anie.202218783