Activity-based protein profiling of serine hydrolases and penicillin-binding proteins in Enterococcus faecium .

Autor:	Grunnvåg JS; Research Group for Host-Microbe Interactions, Department of Medical Biology, UiT - The Arctic University of Norway, Postboks 6050 Langnes, 9037 Tromsø, Norway.; Centre for New Antibacterial Strategies (CANS), UiT - The Arctic University of Norway, Postboks 6050 Langnes, 9037 Tromsø, Norway., Hegstad K; Research Group for Host-Microbe Interactions, Department of Medical Biology, UiT - The Arctic University of Norway, Postboks 6050 Langnes, 9037 Tromsø, Norway.; Centre for New Antibacterial Strategies (CANS), UiT - The Arctic University of Norway, Postboks 6050 Langnes, 9037 Tromsø, Norway.; Norwegian National Advisory Unit on Detection of Antimicrobial Resistance, Department of Microbiology and Infection Control, University Hospital of North Norway, P.O. Box 56, 9038 Tromsø, Norway., Lentz CS; Research Group for Host-Microbe Interactions, Department of Medical Biology, UiT - The Arctic University of Norway, Postboks 6050 Langnes, 9037 Tromsø, Norway.; Centre for New Antibacterial Strategies (CANS), UiT - The Arctic University of Norway, Postboks 6050 Langnes, 9037 Tromsø, Norway.
Jazyk:	angličtina
Zdroj:	FEMS microbes [FEMS Microbes] 2024 May 15; Vol. 5, pp. xtae015. Date of Electronic Publication: 2024 May 15 (Print Publication: 2024).
DOI:	10.1093/femsmc/xtae015
Abstrakt:	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. Competing Interests: None declared. (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)
Databáze:	MEDLINE
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