Activity-based protein profiling of serine hydrolases and penicillin-binding proteins in Enterococcus faecium
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https://hdl.handle.net/10037/34800Dato
2024-05-15Type
Journal articleTidsskriftartikkel
Peer reviewed
Sammendrag
Enterococcus faecium is a gut commensal bacterium which is gaining increasing relevance as an opportunistic, nosocomial pathogen.
Its high level of intrinsic and acquired antimicrobial resistance is causing a lack of treatment options, particularly for infections with
vancomycin-resistant strains, and prioritizes the identification and functional validation of novel druggable targets. Here, we use
activity-based protein profiling (ABPP), a chemoproteomics approach using functionalized covalent inhibitors, to detect active serine
hydrolases across 11 E. faecium and Enterococcus lactis strains. Serine hydrolases are a big and diverse enzyme family, that includes
known drug targets such as penicillin-binding proteins (PBPs), whereas other subfamilies are underexplored. Comparative gel-based
ABPP using Bocillin-FL revealed strain- and growth condition-dependent variations in PBP activities. Profiling with the broadly serine hydrolase-reactive fluorescent probe fluorophosphonate-TMR showed a high similarity across E. faecium clade A1 strains, but
higher variation across A2 and E. lactis strains. To identify these serine hydrolases, we used a biotinylated probe analog allowing
for enrichment and identification via liquid chromatography–mass spectrometry. We identified 11 largely uncharacterized targets
(α,β-hydrolases, SGNH-hydrolases, phospholipases, and amidases, peptidases) that are druggable and accessible in live vancomycinresistant E. faecium E745 and may possess vital functions that are to be characterized in future studies.
Forlag
Oxford University PressSitering
Grunnvåg, Hegstad, Lentz. Activity-based protein profiling of serine hydrolases and penicillin-binding proteins in Enterococcus faecium. FEMS microbes. 2024;5Metadata
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