dc.contributor.author | Uddin, Md Jalal | |
dc.contributor.author | Overkleeft, Hermen S. | |
dc.contributor.author | Lentz, Christian Stephan | |
dc.date.accessioned | 2023-11-21T09:43:14Z | |
dc.date.available | 2023-11-21T09:43:14Z | |
dc.date.issued | 2023-08-08 | |
dc.description.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 | en_US |
dc.identifier.citation | Uddin, Overkleeft, Lentz. Activity-Based Protein Profiling in Methicillin-Resistant Staphylococcus aureus Reveals the Broad Reactivity of a Carmofur-Derived Probe. ChemBioChem. 2023 | en_US |
dc.identifier.cristinID | FRIDAID 2185937 | |
dc.identifier.doi | 10.1002/cbic.202300473 | |
dc.identifier.issn | 1439-4227 | |
dc.identifier.issn | 1439-7633 | |
dc.identifier.uri | https://hdl.handle.net/10037/31831 | |
dc.language.iso | eng | en_US |
dc.publisher | Wiley | en_US |
dc.relation.ispartof | Uddin, 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.journal | ChemBioChem | |
dc.rights.accessRights | openAccess | en_US |
dc.rights.holder | Copyright 2023 The Author(s) | en_US |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0 | en_US |
dc.rights | Attribution 4.0 International (CC BY 4.0) | en_US |
dc.title | Activity-Based Protein Profiling in Methicillin-Resistant Staphylococcus aureus Reveals the Broad Reactivity of a Carmofur-Derived Probe | en_US |
dc.type.version | publishedVersion | en_US |
dc.type | Journal article | en_US |
dc.type | Tidsskriftartikkel | en_US |
dc.type | Peer reviewed | en_US |