PXD062775-1

PXD062775 is an original dataset announced via ProteomeXchange.

Title	Identification of receptor-binding domains of Bacteroidales antibacterial pore-forming toxins
Description	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 demonstrate that receptor recognition is entirely driven by the C-terminal domain (CTD) of BSAP-1 using a combination of in vivo competition assays, in vitro protein binding studies and structural analysis. 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.
HostingRepository	PRIDE
AnnounceDate	2025-11-29
AnnouncementXML	Submission_2025-11-29_00:49:46.981.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD062775
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Frederic Lauber
SpeciesList	scientific name: Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482); NCBI TaxID: NEWT:226186;
ModificationList	2-pyrrolidone-5-carboxylic acid (Glu); 2-pyrrolidone-5-carboxylic acid (Gln); alkylated residue; monohydroxylated residue
Instrument	Orbitrap Exploris 240

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-09 01:20:53	ID requested
⏵ 1	2025-11-29 00:49:47	announced

10.6019/PXD062775;

submitter keyword: LC-MSMS,Bacteroides fragilis, BSAP receptor, co-purification, membrane fraction

Frederic Lauber
contact affiliation	De Duve Institute - University of Louvain
contact email	frederic.lauber@uclouvain.be
lab head
Frederic Lauber
contact affiliation	De Duve Institute - University of Louvain
contact email	frederic.lauber@uclouvain.be
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD062775
     1. Label: PRIDE project
     2. Name: Identification of receptor-binding domains of Bacteroidales antibacterial pore-forming toxins