Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell
Abstract Many proteins of the Repeats in Toxins (RTX) protein family are toxins of Gram-negative pathogens including hemolysin A (HlyA) of uropathogenic E. coli. RTX proteins are secreted via Type I secretion systems (T1SS) and adopt their native conformation in the Ca2+-rich extracellular environme...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2022-10-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-22480-x |
| _version_ | 1828731731925008384 |
|---|---|
| author | Isabelle N. Erenburg Sebastian Hänsch Feby M. Chacko Anna Hamacher Sebastian Wintgens Fabian Stuhldreier Gereon Poschmann Olivia Spitz Kai Stühler Sebastian Wesselborg Johannes H. Hegemann Sander H. J. Smits Stefanie Weidtkamp-Peters Lutz Schmitt |
| author_facet | Isabelle N. Erenburg Sebastian Hänsch Feby M. Chacko Anna Hamacher Sebastian Wintgens Fabian Stuhldreier Gereon Poschmann Olivia Spitz Kai Stühler Sebastian Wesselborg Johannes H. Hegemann Sander H. J. Smits Stefanie Weidtkamp-Peters Lutz Schmitt |
| author_sort | Isabelle N. Erenburg |
| collection | DOAJ |
| description | Abstract Many proteins of the Repeats in Toxins (RTX) protein family are toxins of Gram-negative pathogens including hemolysin A (HlyA) of uropathogenic E. coli. RTX proteins are secreted via Type I secretion systems (T1SS) and adopt their native conformation in the Ca2+-rich extracellular environment. Here we employed the E. coli HlyA T1SS as a heterologous surrogate system for the RTX toxin MbxA from the bovine pathogen Moraxella bovis. In E. coli the HlyA system successfully activates the heterologous MbxA substrate by acylation and secretes the precursor proMbxA and active MbxA allowing purification of both species in quantities sufficient for a variety of investigations. The activating E. coli acyltransferase HlyC recognizes the acylation sites in MbxA, but unexpectedly in a different acylation pattern as for its endogenous substrate HlyA. HlyC-activated MbxA shows host species-independent activity including a so-far unknown toxicity against human lymphocytes and epithelial cells. Using live-cell imaging, we show an immediate MbxA-mediated permeabilization and a rapidly developing blebbing of the plasma membrane in epithelial cells, which is associated with immediate cell death. |
| first_indexed | 2024-04-12T17:50:43Z |
| format | Article |
| id | doaj.art-37bf837447c345038a8921ad00a7000f |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-12T17:50:43Z |
| publishDate | 2022-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-37bf837447c345038a8921ad00a7000f2022-12-22T03:22:31ZengNature PortfolioScientific Reports2045-23222022-10-0112111810.1038/s41598-022-22480-xHeterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cellIsabelle N. Erenburg0Sebastian Hänsch1Feby M. Chacko2Anna Hamacher3Sebastian Wintgens4Fabian Stuhldreier5Gereon Poschmann6Olivia Spitz7Kai Stühler8Sebastian Wesselborg9Johannes H. Hegemann10Sander H. J. Smits11Stefanie Weidtkamp-Peters12Lutz Schmitt13Institute of Biochemistry, Heinrich-Heine-University DüsseldorfCenter for Advanced Imaging, Heinrich Heine University DüsseldorfInstitute of Biochemistry, Heinrich-Heine-University DüsseldorfCenter for Advanced Imaging, Heinrich Heine University DüsseldorfInstitute of Functional Microbial Genomics, Heinrich Heine University DüsseldorfInstitute of Molecular Medicine I, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University DüsseldorfInstitute of Molecular Medicine, Protein Research, University Hospital and Medical Faculty, Heinrich Heine University DüsseldorfInstitute of Biochemistry, Heinrich-Heine-University DüsseldorfInstitute of Molecular Medicine, Protein Research, University Hospital and Medical Faculty, Heinrich Heine University DüsseldorfInstitute of Molecular Medicine I, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University DüsseldorfInstitute of Functional Microbial Genomics, Heinrich Heine University DüsseldorfInstitute of Biochemistry, Heinrich-Heine-University DüsseldorfCenter for Advanced Imaging, Heinrich Heine University DüsseldorfInstitute of Biochemistry, Heinrich-Heine-University DüsseldorfAbstract Many proteins of the Repeats in Toxins (RTX) protein family are toxins of Gram-negative pathogens including hemolysin A (HlyA) of uropathogenic E. coli. RTX proteins are secreted via Type I secretion systems (T1SS) and adopt their native conformation in the Ca2+-rich extracellular environment. Here we employed the E. coli HlyA T1SS as a heterologous surrogate system for the RTX toxin MbxA from the bovine pathogen Moraxella bovis. In E. coli the HlyA system successfully activates the heterologous MbxA substrate by acylation and secretes the precursor proMbxA and active MbxA allowing purification of both species in quantities sufficient for a variety of investigations. The activating E. coli acyltransferase HlyC recognizes the acylation sites in MbxA, but unexpectedly in a different acylation pattern as for its endogenous substrate HlyA. HlyC-activated MbxA shows host species-independent activity including a so-far unknown toxicity against human lymphocytes and epithelial cells. Using live-cell imaging, we show an immediate MbxA-mediated permeabilization and a rapidly developing blebbing of the plasma membrane in epithelial cells, which is associated with immediate cell death.https://doi.org/10.1038/s41598-022-22480-x |
| spellingShingle | Isabelle N. Erenburg Sebastian Hänsch Feby M. Chacko Anna Hamacher Sebastian Wintgens Fabian Stuhldreier Gereon Poschmann Olivia Spitz Kai Stühler Sebastian Wesselborg Johannes H. Hegemann Sander H. J. Smits Stefanie Weidtkamp-Peters Lutz Schmitt Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell Scientific Reports |
| title | Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell |
| title_full | Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell |
| title_fullStr | Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell |
| title_full_unstemmed | Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell |
| title_short | Heterologously secreted MbxA from Moraxella bovis induces a membrane blebbing response of the human host cell |
| title_sort | heterologously secreted mbxa from moraxella bovis induces a membrane blebbing response of the human host cell |
| url | https://doi.org/10.1038/s41598-022-22480-x |
| work_keys_str_mv | AT isabellenerenburg heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT sebastianhansch heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT febymchacko heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT annahamacher heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT sebastianwintgens heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT fabianstuhldreier heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT gereonposchmann heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT oliviaspitz heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT kaistuhler heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT sebastianwesselborg heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT johanneshhegemann heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT sanderhjsmits heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT stefanieweidtkamppeters heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell AT lutzschmitt heterologouslysecretedmbxafrommoraxellabovisinducesamembraneblebbingresponseofthehumanhostcell |