Darobactin B stabilises a lateral-closed conformation of the BAM complex in E. coli cells

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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Downloads (Pure)


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.
Original languageEnglish
Article numbere202218783
Number of pages8
JournalAngewandte Chemie International Edition
Issue number34
Early online date31 May 2023
Publication statusPublished - 21 Aug 2023


  • BAM
  • Darobactin
  • In-cell EPR
  • CryoEM


Dive into the research topics of 'Darobactin B stabilises a lateral-closed conformation of the BAM complex in E. coli cells'. Together they form a unique fingerprint.

Cite this