Abstract
The bacterial type IV pilus (T4P) is a prominent virulence factor in many significant human pathogens, some of which have become increasingly antibiotic resistant. Antivirulence chemotherapeutics are considered a promising alternative to antibiotics because they target the disease process instead of bacterial viability. However, a roadblock to the discovery of anti-T4P compounds is the lack of a high throughput screen (HTS) that can be implemented relatively easily and economically. Here, we describe the first HTS for the identification of inhibitors specifically against the T4P assembly ATPase PilB in vitro. Chloracidobacterium thermophilum PilB (CtPilB) had been demonstrated to have robust ATPase activity and the ability to bind its expected ligands in vitro. We utilized CtPilB and MANT-ATP, a fluorescent ATP analog, to develop a binding assay and adapted it for an HTS. As a proof of principle, we performed a pilot screen with a small compound library of kinase inhibitors and identified quercetin as a PilB inhibitor in vitro. Using Myxococcus xanthus as a model bacterium, we found quercetin to reduce its T4P-dependent motility and T4P assembly in vivo. These results validated our HTS as effective in identifying PilB inhibitors. This assay may prove valuable in seeking leads for the development of antivirulence chemotherapeutics against PilB, an essential and universal component of all bacterial T4P systems.
Recommended Citation
K. J. Dye et al., "High-Throughput Screen for Inhibitors of the Type IV Pilus Assembly ATPase PilB," mSphere, vol. 6, no. 2, pp. 1 - 12, American Society for Microbiology, Mar 2021.
The definitive version is available at https://doi.org/10.1128/mSphere.00129-21
Department(s)
Chemistry
Publication Status
Open Access
Keywords and Phrases
antivirulence; high-throughput screen (HTS); Myxococcus xanthus; PilB ATPase; quercetin, motility; type IV pili (T4P)
International Standard Serial Number (ISSN)
2379-5042
Document Type
Article - Journal
Document Version
Final Version
File Type
text
Language(s)
English
Rights
© 2024 The Authors, All rights reserved.
Creative Commons Licensing
This work is licensed under a Creative Commons Attribution 4.0 License.
Publication Date
01 Mar 2021
PubMed ID
33658276
Comments
National Science Foundation, Grant 1417726