PXD060170

PXD060170 is an original dataset announced via ProteomeXchange.

Title	Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus
Description	Enteric bacterial pathogens pose significant threats to human health; however, the mechanisms by which they infect the mammalian gut in the face of daunting host defenses remain to be fully defined. For the attaching and effacing (A/E) bacterial family member and murine pathogen Citrobacter rodentium, its virulence strategy appears to involve penetration of the colonic mucus barrier, to reach the underlying epithelium. To better define these interactions, we grew colonoids under air-liquid interface (ALI) conditions, producing a thick mucus layer that mimicked in vivo mucus composition and glycosylation. C. rodentium’s penetration of ALI mucus was dramatically enhanced upon exposure to sialic acid, in concert with the secretion of two autotransporter (SPATE) proteins, Pic and EspC. Despite Pic being a class II SPATE, and already recognized as a mucinase, it was EspC, belonging to the class I SPATE family, and not previously shown to possess mucinase activity, that degraded the ALI-derived mucus. Confirming this finding, E. coli DH5 carrying a plasmid that expresses C. rodentium-derived EspC was able to degrade the mucus. Notably, recombinant EspC alone also displayed mucinolytic activity in a dose-dependent manner. Collectively, our results reveal the utility of ALI-derived mucus in modeling microbe-host interactions at the intestinal mucosal surface, as well as identify EspC as an atypical Class I SPATE that shows significant mucinolytic activity towards ALI-derived mucus.
HostingRepository	PRIDE
AnnounceDate	2026-02-03
AnnouncementXML	Submission_2026-02-03_15:08:01.330.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Dominique Levesque
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: NEWT:10090;
ModificationList	iodoacetamide derivatized residue
Instrument	timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2025-01-24 13:16:00	ID requested
⏵ 1	2026-02-03 15:08:01	announced

10.1080/19490976.2025.2494717;

submitter keyword: mucus., EspC,Bacterial pathogens, Air-liquid interface, colonoids

Bruce A. Vallance
contact affiliation	Division of Gastroenterology, Department of Pediatrics, University of British Columbia, Vancouver, BC, Canada.
contact email	bvallance@cw.bc.ca
lab head
Dominique Levesque
contact affiliation	University of Sherbrooke, FM Boisvert lab
contact email	dominique.levesque@usherbrooke.ca
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD060170
     1. Label: PRIDE project
     2. Name: Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus