Application of Fragment-Based Screening to the Design of Inhibitors of Escherichia coli DsbA

• Published in: Angewandte Chemie (International ed.) Vol. 54; no. 7; pp. 2179 - 2184
• Main Authors: Adams, Luke A., Sharma, Pooja, Mohanty, Biswaranjan, Ilyichova, Olga V., Mulcair, Mark D., Williams, Martin L., Gleeson, Ellen C., Totsika, Makrina, Doak, Bradley C., Caria, Sofia, Rimmer, Kieran, Horne, James, Shouldice, Stephen R., Vazirani, Mansha, Headey, Stephen J., Plumb, Brent R., Martin, Jennifer L., Heras, Begoña, Simpson, Jamie S., Scanlon, Martin J.
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 09.02.2015; WILEY‐VCH Verlag; Wiley; Wiley Subscription Services, Inc
• Edition: International ed. in English
• Subjects: Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Bacteria; bacterial virulence; Bacteriology; Chemistry; Chemistry, Multidisciplinary; Crystal structure; Drug Design; E coli; EcDsbA; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Enzymes; Escherichia coli; Escherichia coli - drug effects; Escherichia coli - enzymology; Escherichia coli Infections - drug therapy; Escherichia coli Infections - microbiology; Escherichia coli Proteins - antagonists & inhibitors; Escherichia coli Proteins - metabolism; fragment-based drug discovery; Humans; Inhibitors; medicinal chemistry; Molecular Docking Simulation; Mutation; Physical Sciences; Protein Disulfide-Isomerases - antagonists & inhibitors; Protein Disulfide-Isomerases - metabolism; Proteins; Science & Technology; Screening; Virulence; bacterial virulence; fragment-based drug discovery; TARGETING VIRULENCE; ANTIBIOTICS; COMPLEX; drug design; CRYSTAL-STRUCTURE; DISULFIDE; EcDsbA; medicinal chemistry; REVEALS
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201410341

The thiol‐disulfide oxidoreductase enzyme DsbA catalyzes the formation of disulfide bonds in the periplasm of Gram‐negative bacteria. DsbA substrates include proteins involved in bacterial virulence. In the absence of DsbA, many of these proteins do not fold correctly, which renders the bacteria avirulent. Thus DsbA is a critical mediator of virulence and inhibitors may act as antivirulence agents. Biophysical screening has been employed to identify fragments that bind to DsbA from Escherichia coli. Elaboration of one of these fragments produced compounds that inhibit DsbA activity in vitro. In cell‐based assays, the compounds inhibit bacterial motility, but have no effect on growth in liquid culture, which is consistent with selective inhibition of DsbA. Crystal structures of inhibitors bound to DsbA indicate that they bind adjacent to the active site. Together, the data suggest that DsbA may be amenable to the development of novel antibacterial compounds that act by inhibiting bacterial virulence. Combating bacterial virulence: DsbA is an oxidoreductase enzyme and a key mediator of virulence in Escherichia coli. Using fragment‐based screening, compounds were developed that inhibit DsbA activity in vitro and E. coli motility in a cell‐based assay. Crystal structures of the compounds in complex with DsbA provide a rationale for their activity.