Inhibition of autotransporter biogenesis by small molecules

Maurice Steenhuis, Abdallah M. Abdallah, Sabrina M. de Munnik, Sebastiaan Kuhne, Geert Jan Sterk, Bart van den Berg van Saparoea, Sibel Westerhausen, Samuel Wagner, Nicole N. van der Wel, Maikel Wijtmans, Peter van Ulsen, Wouter S.P. Jong, Joen Luirink

Research output: Contribution to JournalArticleAcademicpeer-review

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.

Original languageEnglish
Pages (from-to)81-98
Number of pages18
JournalMolecular Microbiology
Volume112
Issue number1
DOIs
Publication statusPublished - 1 Jul 2019

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Membrane Proteins
Virulence Factors
Libraries
Hemoglobins
Peptide Hydrolases
Gram-Negative Bacteria
Action Potentials
Fluorescence
Anti-Bacterial Agents
Antigens
Membranes
Type V Secretion Systems
Pharmaceutical Preparations

Cite this

Steenhuis, Maurice ; Abdallah, Abdallah M. ; de Munnik, Sabrina M. ; Kuhne, Sebastiaan ; Sterk, Geert Jan ; van den Berg van Saparoea, Bart ; Westerhausen, Sibel ; Wagner, Samuel ; van der Wel, Nicole N. ; Wijtmans, Maikel ; van Ulsen, Peter ; Jong, Wouter S.P. ; Luirink, Joen. / Inhibition of autotransporter biogenesis by small molecules. In: Molecular Microbiology. 2019 ; Vol. 112, No. 1. pp. 81-98.
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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.",
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Steenhuis, M, Abdallah, AM, de Munnik, SM, Kuhne, S, Sterk, GJ, van den Berg van Saparoea, B, Westerhausen, S, Wagner, S, van der Wel, NN, Wijtmans, M, van Ulsen, P, Jong, WSP & Luirink, J 2019, 'Inhibition of autotransporter biogenesis by small molecules' Molecular Microbiology, vol. 112, no. 1, pp. 81-98. https://doi.org/10.1111/mmi.14255

Inhibition of autotransporter biogenesis by small molecules. / Steenhuis, Maurice; Abdallah, Abdallah M.; de Munnik, Sabrina M.; Kuhne, Sebastiaan; Sterk, Geert Jan; van den Berg van Saparoea, Bart; Westerhausen, Sibel; Wagner, Samuel; van der Wel, Nicole N.; Wijtmans, Maikel; van Ulsen, Peter; Jong, Wouter S.P.; Luirink, Joen.

In: Molecular Microbiology, Vol. 112, No. 1, 01.07.2019, p. 81-98.

Research output: Contribution to JournalArticleAcademicpeer-review

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Steenhuis M, Abdallah AM, de Munnik SM, Kuhne S, Sterk GJ, van den Berg van Saparoea B et al. Inhibition of autotransporter biogenesis by small molecules. Molecular Microbiology. 2019 Jul 1;112(1):81-98. https://doi.org/10.1111/mmi.14255