PXD039143

PXD039143 is an original dataset announced via ProteomeXchange.

Title	Characterizing metabolic drivers of Clostridioides difficile infection with activity-based hydrazine probes
Description	Many enzymes require post-translational modifications or cofactor machinery for primary function. As these catalytically essential moieties are highly regulated, they act as dual sensors and chemical handles for context-dependent metabolic activity. Clostridioides difficile is a major nosocomial pathogen that infects the colon. Energy generating metabolism, particularly through amino acid Stickland fermentation, is central to colonization and persistence of this pathogen during infection. Here using activity-based protein profiling (ABPP), we revealed Stickland enzyme activity is a biomarker for C. difficile infection (CDI) and annotated two such cofactor-dependent Stickland reductases. We, for the first time, structurally assigned proline reductase and glycine reductase as cysteine-derived pyruvoyl-dependent enzymes and showed through cofactor monitoring that their activity is regulated by their respective amino acid substrates. Proline reductase was consistently active in toxigenic C. difficile, confirming the enzyme to be a major metabolic driver of CDI. Further, activity-based hydrazine probes were shown to be active site directed inhibitors of proline reductase. As such, this enzyme activity, via its druggable cofactor modality, is a promising therapeutic target that could allow for the repopulation of bacteria that compete with C. difficile for proline and therefore restore colonization resistance against C. difficile in the gut.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:43:51.788.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD039143
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Megan Matthews
SpeciesList	scientific name: Clostridioides difficile ATCC 43255; NCBI TaxID: 499175;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2023-01-02 01:29:15	ID requested
1	2023-03-11 11:20:00	announced
⏵ 2	2023-11-14 08:43:59	announced	2023-11-14: Updated project metadata.

10.6019/PXD039143;

Bustin KA, Abbas A, Wang X, Abt MC, Zackular JP, Matthews ML, infection with activity-based hydrazine probes. Front Pharmacol, 14():1074619(2023) [pubmed]

submitter keyword: metabolism, C. difficile, cofactor, hydrazine,ABPP

Megan L. Matthews
contact affiliation	University of Pennsylvania
contact email	megamatt@sas.upenn.edu
lab head
Megan Matthews
contact affiliation	University of Pennsylvania
contact email	megamatt@sas.upenn.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/03/PXD039143

PRIDE project URI

• PRIDE
  1. PXD039143
     1. Label: PRIDE project
     2. Name: Characterizing metabolic drivers of Clostridioides difficile infection with activity-based hydrazine probes