L-type pyocins inhibit the BAM complex to kill without cell entry

Fabian Munder; Matthew D Johnson; Imogen Samuels; Laura McCaughey; Oleksii Zdorevskyi; Chunxiao Wang; Ashleigh Kropp; Lauren Zavan; Erin P Price; Derek S Sarovich; Swati Varshney; Christopher A McDevitt; Hari Venugopal; Vivek Sharma; Matthew T Doyle; Francesca Short; Debnath Ghosal; James P R Connolly; Gavin J Knott; Rhys Grinter

doi:10.1038/s41467-026-74995-w

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L-type pyocins inhibit the BAM complex to kill without cell entry

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L-type pyocins inhibit the BAM complex to kill without cell entry

Fabian Munder, Matthew D Johnson, Imogen Samuels, Laura McCaughey, Oleksii Zdorevskyi, Chunxiao Wang, Ashleigh Kropp, Lauren Zavan, Erin P Price, Derek S Sarovich, …

Nature Communications, Vol.Advanced access

02-Jul-2026

DOI: https://doi.org/10.1038/s41467-026-74995-w

PMID: 42393045

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Abstract

antibiotics

bacteriology

cryoelectron microscopy

target identification

Many antibiotics are ineffective against the Gram-negative pathogen Pseudomonas aeruginosa because of intrinsic defence mechanisms, such as the impermeable bacterial outer membrane. Here, we show that protein antibiotics called L-type pyocins kill P. aeruginosa by inhibiting the β-barrel assembly machinery (BAM) complex at the cell surface, halting outer-membrane protein assembly. Using single-particle cryo-electron microscopy, we show that L-type pyocins bind a surface-exposed region of BamA and deploy a C-terminal peptide that competitively inhibits the BAM complex, demonstrating that cell entry is not required for antibiotic activity. We combine genetics, multi-omics and cryo-electron tomography to show that BAM complex inhibition by L-type pyocins or the cyclic-peptide antibiotic, darobactin, triggers a multifaceted transcriptomic, proteomic, and morphological response. BAM inhibition ultimately leads to a catastrophic loss of membrane integrity and cell death. These results validate BAM as a target for antibiotics that do not enter the cell and define an engineerable system for their development.

Details

Title: L-type pyocins inhibit the BAM complex to kill without cell entry
Authors: Fabian Munder - The University of Melbourne
Matthew D Johnson - The University of Melbourne
Imogen Samuels - The University of Melbourne
Laura McCaughey - University of Glasgow
Oleksii Zdorevskyi - University of Helsinki
Chunxiao Wang - The University of Melbourne
Ashleigh Kropp - The University of Melbourne
Lauren Zavan - The University of Melbourne
Erin P Price - University of the Sunshine Coast
Derek S Sarovich - University of the Sunshine Coast
Swati Varshney - The University of Melbourne
Christopher A McDevitt - The University of Melbourne
Hari Venugopal - Monash University
Vivek Sharma - University of Helsinki
Matthew T Doyle - The University of Sydney
Francesca Short - Monash University
Debnath Ghosal - The University of Melbourne
James P R Connolly - Newcastle University
Gavin J Knott - Monash University
Rhys Grinter (Corresponding Author) - The University of Melbourne
Publication details: Nature Communications, Vol.Advanced access
Publisher: Nature Publishing Group
DOI: 10.1038/s41467-026-74995-w
ISSN: 2041-1723
PMID: 42393045
Copyright note: This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
Grants: Structural role of the host cytoskeleton during invasion of intracellular pathogens, 1196924, National Health and Medical Research Council (Australia, Canberra) - NHMRC
Targeting iron piracy from host proteins by Neisseria and Haemophilus spp. for the development of novel antimicrobials, 1197376, National Health and Medical Research Council (Australia, Canberra) - NHMRC
Harnessing structural insights to block pneumococcal manganese uptake, 2010400, National Health and Medical Research Council (Australia, Canberra) - NHMRC
Taking Control: Understanding regulation of bacterial iron acquisition, DE200101524, Australian Research Council (Australia, Canberra) - ARC
Elucidating the determinants of cation import across the kingdoms of life, DP220100713, Australian Research Council (Australia, Canberra) - ARC
ARC Training Centre for Cryo-Electron Microscopy of Membrane Proteins for Drug Discovery, IC200100052, Australian Research Council (Australia, Canberra) - ARC
Hitting bacteria with a Bam: Lectin-Like Antimicrobials as New Antibiotics, DP230102150, Australian Research Council (Australia, Canberra) - ARC
New generation direct electron detector for cryo-electron microscopy, LE200100045, Australian Research Council (Australia, Canberra) - ARC
A centre for structural cryo-electron microscopy, LE120100090, Australian Research Council (Australia, Canberra) - ARC
Grant note: Snow Medical Research Foundation (SMRF2021-276) / Sir Henry Wellcome Fellowship from the Wellcome Trust 9106064/Z/14/29 / Cumming Global Centre for Pandemic Therapeutics Foundation grant, Peter Doherty Institute for Infection and Immunity (CGCPT-CGCPT00060) / Sir Henry Wellcome Fellowship (106063/A/14/Z) / UKRI Medical Research Council (MR/X007197/1) and Biotechnology and Biological Sciences Research Council (UKRI797) / Wellcome (206194) / Research Council of Finland (362411 / Research Council of Finland (338176), the Sigrid Jusélius Foundation and the Jane and Aatos Erkko Foundation.
Organisation Unit: School of Science, Technology and Engineering; Centre for Bioinnovation
Language: English
Record Identifier: 991242594302621
Output Type: Journal article

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