The discovery and structural basis of two distinct state-dependent inhibitors of BamA
BamA is the central component of the essential β-barrel assembly machine (BAM), a conserved multi-subunit complex that dynamically inserts and folds β-barrel proteins into the outer membrane of Gram-negative bacteria. Despite recent advances in our mechanistic and structural understanding of BamA, there are few potent and selective tool molecules that can bind to and modulate BamA activity. Here, we explored in vitro selection methods and different BamA/BAM protein formulations to discover peptide macrocycles that kill Escherichia coli by targeting extreme conformational states of BamA. Our studies show that Peptide Targeting BamA-1 (PTB1) targets an extracellular divalent cation-dependent binding site and locks BamA into a closed lateral gate conformation. By contrast, PTB2 targets a luminal binding site and traps BamA into an open lateral gate conformation. Our results will inform future antibiotic discovery efforts targeting BamA and provide a template to prospectively discover modulators of other dynamic integral membrane proteins.
Authors
- Man‐Wah Tan (ORCID: https://orcid.org/0000-0002-7748-212X)
- Christopher E. Heise
- Benjamin D. Sellers (ORCID: https://orcid.org/0000-0002-4194-1240)
- Wendy Sandoval (ORCID: https://orcid.org/0000-0002-4672-0762)
- Hany S. Girgis
- Dawei Sun (ORCID: https://orcid.org/0000-0003-1551-4349)
- Dimitry Tegunov (ORCID: https://orcid.org/0000-0001-7019-3221)
- Patrick Reid
- Hiroshi Ijiri
- Stephanie Shriver
- John G. Quinn (ORCID: https://orcid.org/0000-0002-4664-6232)
- Christian N. Cunningham (ORCID: https://orcid.org/0000-0003-3993-660X)
- Wayne J. Fairbrother (ORCID: https://orcid.org/0000-0003-0480-5124)
- Jian Payandeh (ORCID: https://orcid.org/0000-0002-4015-9716)
- Tatsuya Sano
- Steven T. Rutherford (ORCID: https://orcid.org/0000-0002-4758-4248)
- Austin K. Murchison
- Claudio Ciferri (ORCID: https://orcid.org/0000-0002-0804-2411)
- Nicholas J. Skelton (ORCID: https://orcid.org/0009-0004-4024-3805)
- Weijing Liu (ORCID: https://orcid.org/0000-0003-2322-0500)
- Christopher P. Arthur (ORCID: https://orcid.org/0000-0001-9647-1509)
- Matthew C. Johnson (ORCID: https://orcid.org/0000-0002-1477-7801)
- Hayato Yanagida
- Hiroko Inaba
- Ying Yang (ORCID: https://orcid.org/0000-0002-2602-6191)
- Kelly M. Storek (ORCID: https://orcid.org/0000-0002-2289-0214)
- Guanghui Han (ORCID: https://orcid.org/0000-0002-9893-9168)
- Mary Kate Alexander (ORCID: https://orcid.org/0009-0004-2041-5016)
- Peter A. Smith (ORCID: https://orcid.org/0000-0002-6631-0613)
- Christine Tam
- Jun‐ichi Nishikawa
Institutions
- GenVec (US)
- Southeastern University (US)
- Exelixis (United States) (US)
- Alameda Hospital (US)
- TCI Peptide Therapeutics (United States) (US)
- Helix (United States) (US)
- Peptidream (Japan) (JP)
- Altos Labs
- Sunesis (United States) (US)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-10-08
- DOI
- https://doi.org/10.1038/s41467-024-52512-1
- Citations
- 19
- Primary Topic
- RNA and protein synthesis mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 4.44
Funders
- Genentech