8BVQ

E. coli BAM complex (BamABCDE) bound to darobactin B

Summary for 8BVQ

• Entry DOI: 10.2210/pdb8bvq/pdb
• EMDB information: 16268
• Related PRD ID: PRD_002342
• Descriptor: Outer membrane protein assembly factor BamA, Outer membrane protein assembly factor BamB, Outer membrane protein assembly factor BamC, ... (6 entities in total)
• Functional Keywords: outer membrane protein, protein folding, antibiotic, beta barrel, membrane protein
• Biological source: Escherichia coli; More
• Total number of polymer chains: 6
• Total formula weight: 201281.21

Authors

Horne, J.E.,Fenn, K.L.,Radford, S.E.,Ranson, N.A. (deposition date: 2022-12-05, release date: 2023-05-24, Last modification date: 2023-09-06)

Primary citation

Haysom, S.F.,Machin, J.,Whitehouse, J.M.,Horne, J.E.,Fenn, K.,Ma, Y.,El Mkami, H.,Bohringer, N.,Schaberle, T.F.,Ranson, N.A.,Radford, S.E.,Pliotas, C.
Darobactin B Stabilises a Lateral-Closed Conformation of the BAM Complex in E. coli Cells.
Angew.Chem.Int.Ed.Engl., 62:e202218783-e202218783, 2023

PubMed 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.

PubMed: 37162386
DOI: 10.1002/anie.202218783

Experimental method

ELECTRON MICROSCOPY (3.3 Å)