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Inhibition of autotransporter biogenesis by small molecules
Author(s) -
Steenhuis Maurice,
Abdallah Abdallah M.,
de Munnik Sabrina M.,
Kuhne Sebastiaan,
Sterk GeertJan,
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
Publication year - 2019
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.14255
Subject(s) - secretion , biology , biogenesis , bacterial outer membrane , virulence , microbiology and biotechnology , cell envelope , secretory protein , vesicle , membrane protein , mutant , biochemistry , membrane , escherichia coli , gene
Summary 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.