Activity-Based Protein Profiling in Methicillin-Resistant Staphylococcus aureus Reveals the Broad Reactivity of a Carmofur-Derived Probe
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https://hdl.handle.net/10037/31831Date
2023-08-08Type
Journal articleTidsskriftartikkel
Peer reviewed
Abstract
Activity-based protein profiling is a powerful chemoproteomic
technique to detect active enzymes and identify targets and
off-targets of drugs. Here, we report the use of carmofur- and
activity-based probes to identify biologically relevant enzymes
in the bacterial pathogen Staphylococcus aureus. Carmofur is an
anti-neoplastic prodrug of 5-fluorouracil and also has antimicrobial and anti-biofilm activity. Carmofur probes were originally
designed to target human acid ceramidase, a member of the
NTN hydrolase family with an active-site cysteine nucleophile.
Here, we first profiled the targets of a fluorescent carmofur
probe in live S. aureus under biofilm-promoting conditions and
in liquid culture, before proceeding to target identification by
liquid chromatography/mass spectrometry. Treatment with a
carmofur-biotin probe led to enrichment of 20 enzymes from
diverse families awaiting further characterization, including the
NTN hydrolase-related IMP cyclohydrolase PurH. However, the
probe preferentially labeled serine hydrolases, thus displaying a
reactivity profile similar to that of carbamates. Our results
suggest that the electrophilic N-carbamoyl-5-fluorouracil scaffold could potentially be optimized to achieve selectivity
towards diverse enzyme families. The observed promiscuous
reactivity profile suggests that the clinical use of carmofur
presumably leads to inactivation of a number human and
microbial enzymes, which could lead to side effects and/or
contribute to therapeutic efficacy
Is part of
Uddin, M.J. (2024). Activity-Based Protein Profiling (ABPP) and functional validation of uncharacterized enzymes in bacterial pathogens Staphylococcus aureus and Klebsiella pneumoniae. (Doctoral thesis). https://hdl.handle.net/10037/34285Publisher
WileyCitation
Uddin, Overkleeft, Lentz. Activity-Based Protein Profiling in Methicillin-Resistant Staphylococcus aureus Reveals the Broad Reactivity of a Carmofur-Derived Probe. ChemBioChem. 2023Metadata
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