Inhibition of autotransporter biogenesis by small molecules

M. Steenhuis, S. Luirink, W.S.P. Jong, J.P. van Ulsen, Maikel Wijtmans, H.B. van den Berg van Saparoea, G.J. Sterk, S. Kuhne, S.M. de Munnik, Abdallah M. Abdallah, N.N. van der Wel, Samuel Wagner, Sibel Westerhausen

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)1-18
Number of pages18
JournalMolecular Microbiology
Publication statusPublished - 28 Mar 2019

Fingerprint

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, M. ; Luirink, S. ; Jong, W.S.P. ; van Ulsen, J.P. ; Wijtmans, Maikel ; van den Berg van Saparoea, H.B. ; Sterk, G.J. ; Kuhne, S. ; de Munnik, S.M. ; Abdallah, Abdallah M. ; van der Wel, N.N. ; Wagner, Samuel ; Westerhausen, Sibel. / Inhibition of autotransporter biogenesis by small molecules. In: Molecular Microbiology. 2019 ; pp. 1-18.
@article{44577a99e30a40718b1899c21eb19bf1,
title = "Inhibition of autotransporter biogenesis by small molecules",
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 assayincites its use to screen larger compound libraries with drug-like compounds.",
author = "M. Steenhuis and S. Luirink and W.S.P. Jong and {van Ulsen}, J.P. and Maikel Wijtmans and {van den Berg van Saparoea}, H.B. and G.J. Sterk and S. Kuhne and {de Munnik}, S.M. and Abdallah, {Abdallah M.} and {van der Wel}, N.N. and Samuel Wagner and Sibel Westerhausen",
year = "2019",
month = "3",
day = "28",
language = "English",
pages = "1--18",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",

}

Steenhuis, M, Luirink, S, Jong, WSP, van Ulsen, JP, Wijtmans, M, van den Berg van Saparoea, HB, Sterk, GJ, Kuhne, S, de Munnik, SM, Abdallah, AM, van der Wel, NN, Wagner, S & Westerhausen, S 2019, 'Inhibition of autotransporter biogenesis by small molecules' Molecular Microbiology, pp. 1-18.

Inhibition of autotransporter biogenesis by small molecules. / Steenhuis, M.; Luirink, S.; Jong, W.S.P.; van Ulsen, J.P.; Wijtmans, Maikel; van den Berg van Saparoea, H.B.; Sterk, G.J.; Kuhne, S.; de Munnik, S.M.; Abdallah, Abdallah M.; van der Wel, N.N.; Wagner, Samuel; Westerhausen, Sibel.

In: Molecular Microbiology, 28.03.2019, p. 1-18.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Inhibition of autotransporter biogenesis by small molecules

AU - Steenhuis, M.

AU - Luirink, S.

AU - Jong, W.S.P.

AU - van Ulsen, J.P.

AU - Wijtmans, Maikel

AU - van den Berg van Saparoea, H.B.

AU - Sterk, G.J.

AU - Kuhne, S.

AU - de Munnik, S.M.

AU - Abdallah, Abdallah M.

AU - van der Wel, N.N.

AU - Wagner, Samuel

AU - Westerhausen, Sibel

PY - 2019/3/28

Y1 - 2019/3/28

N2 - 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 assayincites its use to screen larger compound libraries with drug-like compounds.

AB - 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 assayincites its use to screen larger compound libraries with drug-like compounds.

UR - https://onlinelibrary.wiley.com/doi/full/10.1111/mmi.14255

M3 - Article

SP - 1

EP - 18

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

ER -