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dc.contributor.authorUddin, Md Jalal
dc.contributor.authorOverkleeft, Hermen S.
dc.contributor.authorLentz, Christian Stephan
dc.date.accessioned2023-11-21T09:43:14Z
dc.date.available2023-11-21T09:43:14Z
dc.date.issued2023-08-08
dc.description.abstractActivity-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 efficacyen_US
dc.identifier.citationUddin, Overkleeft, Lentz. Activity-Based Protein Profiling in Methicillin-Resistant Staphylococcus aureus Reveals the Broad Reactivity of a Carmofur-Derived Probe. ChemBioChem. 2023en_US
dc.identifier.cristinIDFRIDAID 2185937
dc.identifier.doi10.1002/cbic.202300473
dc.identifier.issn1439-4227
dc.identifier.issn1439-7633
dc.identifier.urihttps://hdl.handle.net/10037/31831
dc.language.isoengen_US
dc.publisherWileyen_US
dc.relation.ispartofUddin, M.J. (2024). Activity-Based Protein Profiling (ABPP) and functional validation of uncharacterized enzymes in bacterial pathogens <i>Staphylococcus aureus</i> and <i>Klebsiella pneumoniae</i>. (Doctoral thesis). <a href=https://hdl.handle.net/10037/34285>https://hdl.handle.net/10037/34285</a>
dc.relation.journalChemBioChem
dc.rights.accessRightsopenAccessen_US
dc.rights.holderCopyright 2023 The Author(s)en_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0en_US
dc.rightsAttribution 4.0 International (CC BY 4.0)en_US
dc.titleActivity-Based Protein Profiling in Methicillin-Resistant Staphylococcus aureus Reveals the Broad Reactivity of a Carmofur-Derived Probeen_US
dc.type.versionpublishedVersionen_US
dc.typeJournal articleen_US
dc.typeTidsskriftartikkelen_US
dc.typePeer revieweden_US


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Attribution 4.0 International (CC BY 4.0)
Med mindre det står noe annet, er denne innførselens lisens beskrevet som Attribution 4.0 International (CC BY 4.0)