Fluoromethylketone-Fragment Conjugates Designed as Covalent Modifiers of EcDsbA are Atypical Substrates.
Bradley C DoakRebecca L WhitehouseKieran RimmerMartin WilliamsBegoña HerasSofia CariaOlga IlyichovaMansha VaziraniBiswaranjan MohantyJason B HarperMartin J ScanlonJamie S SimpsonPublished in: ChemMedChem (2024)
Disulfide bond protein A (DsbA) is an oxidoreductase enzyme that catalyzes the formation of disulfide bonds in Gram-negative bacteria. In Escherichia coli, DsbA (EcDsbA) is essential for bacterial virulence, thus inhibitors have the potential to act as antivirulence agents. A fragment-based screen was conducted against EcDsbA and herein we describe the development of a series of compounds based on a phenylthiophene hit identified from the screen. A novel thiol reactive and "clickable" ethynylfluoromethylketone was designed for reaction with azide-functionalized fragments to enable rapid and versatile attachment to a range of fragments. The resulting fluoromethylketone conjugates showed selectivity for reaction with the active site thiol of EcDsbA, however unexpectedly, turnover of the covalent adduct was observed. A mechanism for this turnover was investigated and proposed which may have wider ramifications for covalent reactions with dithiol-disulfide oxidoreducatases.
Keyphrases
- escherichia coli
- high throughput
- bone mineral density
- cancer therapy
- biofilm formation
- pseudomonas aeruginosa
- staphylococcus aureus
- electron transfer
- quantum dots
- antimicrobial resistance
- risk assessment
- drug delivery
- protein protein
- atomic force microscopy
- binding protein
- multidrug resistant
- amino acid
- transition metal
- loop mediated isothermal amplification
- high resolution
- structural basis
- tandem mass spectrometry