Abstract
The rapid rise of antibiotic-resistant bacteria is one of the major concerns in modern medicine. Therefore, to treat bacterial infections, there is an urgent need for new antibacterials–preferably directed against alternative bacterial targets. One such potential target is the preprotein translocation motor SecA. SecA is a peripheral membrane ATPase and a key component of the Sec secretion pathway, the major route for bacterial protein export across or into the cytoplasmic membrane. As SecA is essential for bacterial viability, ubiquitous and highly conserved in bacteria, but not present in eukaryotic cells, it represents an attractive antibacterial target. Using an in silico approach, we have defined several potentially druggable and conserved pockets on the surface of SecA. We show that three of these potentially druggable sites are important for SecA function. A starting collection of ~500 000 commercially available small-molecules was virtually screened against a predicted druggable pocket in the preprotein-binding domain of Escherichia coli SecA using a multi-step virtual ligand screening protocol. The 1040 top-scoring molecules were tested in vitro for inhibition of the translocation ATPase activity of E. coli SecA. Five inhibitors of the translocation ATPase, and not of basal or membrane ATPase, were identified with IC50 values <65 μm. The most potent inhibitor showed an IC50 of 24 μm. The antimicrobial activity was determined for the five most potent SecA inhibitors. Two compounds were found to possess weak antibacterial activity (IC50 ~198 μm) against E. coli, whereas some compounds showed moderate antibacterial activity (IC50 ~100 μm) against Staphylococcus aureus.
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Acknowledgements
We are grateful to T. D’huys for his contribution to the cloning of the saSecA1 Site I mutants; M. Kulharia for technical assistance during the initial phase of the virtual screening process; L. Van Berckelaer, J. Balzarini and D. Schols for cytotoxicity tests; and P. Chaltin and H. Klaassen for useful discussions. This work was supported by grants SBO IWT 50146; FWO (Krediet aan Navorsers) 1500912N and ESCMID Grant 2012; the Cardiovascular Research Institute Maastricht (to GAFN); the Transnational University Limburg (to GAFN) and KUL-Spa (Onderzoekstoelagen 2013; Bijzonder Onderzoeksfonds; KU Leuven; to AE); RiMembR (Vlaanderen Onderzoeksprojecten; #G0C6814N; FWO; to AE); an Excellence grant (#1473 to AE; Greek General Secretariat of Research).
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De Waelheyns, E., Segers, K., Sardis, M. et al. Identification of small-molecule inhibitors against SecA by structure-based virtual ligand screening. J Antibiot 68, 666–673 (2015). https://doi.org/10.1038/ja.2015.53
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DOI: https://doi.org/10.1038/ja.2015.53
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