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Mol Microbiol. 2019 Jul;112(1):81-98. doi: 10.1111/mmi.14255. Epub 2019 May 3.

Inhibition of autotransporter biogenesis by small molecules.

Author information

1
Department of Molecular Microbiology, Amsterdam Institute for Molecules Medicines and Systems (AIMMS), Vrije Universiteit, Amsterdam, the Netherlands.
2
Bioscience Core Laboratory, King Abdullah University of Science and Technology, Thuwal, Jeddah, Kingdom of Saudi Arabia.
3
Department of Chemistry and Pharmaceutical Sciences, Amsterdam Institute for Molecules Medicines and Systems (AIMMS), Vrije Universiteit, Amsterdam, the Netherlands.
4
Interfaculty Institute of Microbiology and Infection Medicine (IMIT), University of Tübingen, Tübingen, Germany.
5
German Center for Infection Research (DZIF), Tübingen, Germany.
6
Department of Medical Biology, Electron Microscopy Center Amsterdam, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.

Abstract

Disarming pathogens by targeting virulence factors is a promising alternative to classic antibiotics. Many virulence factors in Gram-negative bacteria are secreted via the autotransporter (AT) pathway, also known as Type 5 secretion. These factors are secreted with the assistance of two membrane-based protein complexes: Sec and Bam. To identify inhibitors of the AT pathway, we used transcriptomics analysis to develop a fluorescence-based high-throughput assay that reports on the stress induced by the model AT hemoglobin protease (Hbp) when its secretion across the outer membrane is inhibited. Screening a library of 1600 fragments yielded the compound VUF15259 that provokes cell envelope stress and secretion inhibition of the ATs Hbp and Antigen-43. VUF15259 also impairs β-barrel folding activity of various outer membrane proteins. Furthermore, we found that mutants that are compromised in outer membrane protein biogenesis are more susceptible to VUF15259. Finally, VUF15259 induces the release of vesicles that appear to assemble in short chains. Taken together, VUF15259 is the first reported compound that inhibits AT secretion and our data are mostly consistent with VUF15259 interfering with the Bam-complex as potential mode of action. The validation of the presented assay incites its use to screen larger compound libraries with drug-like compounds.

PMID:
30983025
PMCID:
PMC6850105
DOI:
10.1111/mmi.14255

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