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Activity-based protein profiling of serine hydrolases and penicillin-binding proteins in Enterococcus faecium .
- Source :
-
FEMS microbes [FEMS Microbes] 2024 May 15; Vol. 5, pp. xtae015. Date of Electronic Publication: 2024 May 15 (Print Publication: 2024). - Publication Year :
- 2024
-
Abstract
- 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 vancomycin-resistant E. faecium E745 and may possess vital functions that are to be characterized in future studies.<br />Competing Interests: None declared.<br /> (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)
Details
- Language :
- English
- ISSN :
- 2633-6685
- Volume :
- 5
- Database :
- MEDLINE
- Journal :
- FEMS microbes
- Publication Type :
- Academic Journal
- Accession number :
- 38813097
- Full Text :
- https://doi.org/10.1093/femsmc/xtae015