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PreprintCited by 0Antimicrobial Peptides and ActivitiesBacterial Genetics and BiotechnologyAntimicrobial agents and applications

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Identification of receptor-binding domains of Bacteroidales antibacterial pore-forming toxins

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Borgini, Sofia, Iorga, Bogdan, Vertommen, Didier, Collet, Jean-François, Lauber, Frédéric·2025·Journal of Biological Chemistry
DOI 10.1101/2025.01.06.631476PMC PMC12853789

Preprint — published as 10.1016/j.jbc.2025.111113

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Abstract

Bacteroidales are abundant Gram-negative bacteria present in the gut microbiota of most animals, including humans, where they carry out vital functions for host health. To thrive in this competitive environment, Bacteroidales use sophisticated weapons to outmatch competitors. Among these, BSAPs (Bacteroidales Secreted Antimicrobial Proteins) represent a novel class of bactericidal pore-forming toxins that are highly specific to their receptor, typically targeting only a single membrane protein or lipopolysaccharide. The molecular determinants conferring this high selectivity remain unknown. In this study, we therefore investigated the model protein BSAP-1 and determined which of its domains is involved in providing receptor specificity. We clearly demonstrate that receptor recognition is entirely driven by the C-terminal domain (CTD) of BSAP-1 using a combination of in vivo competition assays and in vitro protein binding studies. Specifically, we show that deletion of the CTD abrogates BSAP-1 bactericidal activity by preventing receptor binding, while grafting the CTD to unrelated carrier proteins enables CTD-driven interaction with the BSAP-1 receptor. Building upon this discovery, we show that BSAPs can be categorized according to the structure of their CTD and that BSAPs within the same cluster are likely to target the same type of receptor. Additionally, we show that the CTD of BSAP-1 can be repurposed to generate probes for fluorescent labelling of membrane proteins in live cells. In summary, our research demonstrates that BSAP receptor recognition is driven by their CTD and that these can be engineered to develop novel tools for the investigation of Bacteroidales biology.

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Metadata

Authors
Borgini, S., Iorga, B., Vertommen, D., Collet, J.-F., & Lauber, F.
Affiliations
  • de Duve Institute (BE)
  • Centre National de la Recherche Scientifique (FR)
  • Vrije Universiteit Brussel (BE)
Corresponding author
Lauber, Frédéric
ORCID
Venue
Journal of Biological Chemistry
Publisher
openRxiv
Published
2025-01-06
Volume
302
Issue
2
Pages
111113-111113
ISSN
0021-9258 (ISSN-L)
DOI
10.1101/2025.01.06.631476
PMC
PMC12853789
OpenAlex
W7117476070
Cited by
0
References
1
FWCI
0.00
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • HAL (submittedVersion, other-oa)
  • HAL (submittedVersion, cc-by)
  • Digital Access to Libraries (Université catholique de Louvain (UCL), l'Université de Namur (UNamur) and the Université Saint-Louis (USL-B)) (submittedVersion, other-oa)
  • VUBIR (Vrije Universiteit Brussel) (submittedVersion)
Funders
  • Agence Nationale de la Recherche (ANR-20-PAMR-0010)
  • Fonds De La Recherche Scientifique - FNRS (O008818F, 1B.274.22F, WELBIO-CR-2019C-03)
  • Fédération Wallonie-Bruxelles (17/22‐087, ARC 17/22-087)
  • Fonds Wetenschappelijk Onderzoek (1S60721N\u20131S60723N)
  • Walloon excellence in life sciences and biotechnology
  • National Institutes of Health
  • Université de Namur
  • National Institute of General Medical Sciences (R01GM129325)
  • National Institute of Allergy and Infectious Diseases
Topics
  • Life Sciences › Immunology and Microbiology › Microbiology › Antimicrobial Peptides and Activities
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Immunology and Microbiology › Applied Microbiology and Biotechnology › Antimicrobial agents and applications
Keywords
pore-forming toxins, receptor specificity, C-terminal domain, receptor binding, electrostatic interactions, bactericidal activity, gut microbiota, Gram-negative bacteria
MeSH terms
Protein Domains, Bacterial Proteins, Bacterial Toxins, Humans, Protein Binding, Bacteroidetes, Pore Forming Cytotoxic Proteins

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