A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i>
The opportunistic pathogen <i>Pseudomonas aeruginosa</i> uses quorum sensing to control its virulence. One of its major signal molecules, the <i>Pseudomonas</i> quinolone signal PQS, has high affinity to membranes and is known to be trafficked mainly via outer membrane vesicl...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-11-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/12/11/1656 |
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| author | Alba Arranz San Martín Steffen Lorenz Drees Susanne Fetzner |
| author_facet | Alba Arranz San Martín Steffen Lorenz Drees Susanne Fetzner |
| author_sort | Alba Arranz San Martín |
| collection | DOAJ |
| description | The opportunistic pathogen <i>Pseudomonas aeruginosa</i> uses quorum sensing to control its virulence. One of its major signal molecules, the <i>Pseudomonas</i> quinolone signal PQS, has high affinity to membranes and is known to be trafficked mainly via outer membrane vesicles (OMVs). We previously reported that several 3-hydroxy-4(1<i>H</i>)-quinolone 2,4-dioxygenases (HQDs) catalyze the cleavage of PQS and thus act as quorum quenching enzymes. Further analysis showed that, in contrast to other HQDs, the activity of HQD from <i>Streptomyces bingchenggensis</i> (HQD<i><sub>S</sub>.<sub>b</sub>.</i>) was unexpectedly stabilized by culture supernatants of <i>P. aeruginosa</i>. Interestingly, the stabilizing effect was higher with supernatants from the strain PA14 than with supernatants from the strain PAO1. Heat treatment and lyophilization hardly affected the stabilizing effect; however, fractionation of the supernatant excluded small molecules as stabilizing agents. In a pull-down assay, HQD<i><sub>S</sub>.<sub>b</sub>.</i> appeared to interact with several <i>P. aeruginosa</i> proteins previously found in the OMV proteome. This prompted us to probe the physical interaction of HQD<i><sub>S</sub>.<sub>b</sub>.</i> with prepared extracellular membrane vesicles. Homo-FRET of fluorescently labeled HQD<i><sub>S</sub>.<sub>b</sub>.</i> indeed indicated a spatial clustering of the protein on the vesicles. Binding of a PQS-cleaving enzyme to the OMVs of <i>P. aeruginosa</i> may enhance PQS degradation and is highly reconcilable with its function as a quorum quenching enzyme. |
| first_indexed | 2024-03-09T19:14:46Z |
| format | Article |
| id | doaj.art-fe077a7bba1c4aeba666874fe749ff0c |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-09T19:14:46Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-fe077a7bba1c4aeba666874fe749ff0c2023-11-24T03:54:08ZengMDPI AGBiomolecules2218-273X2022-11-011211165610.3390/biom12111656A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i>Alba Arranz San Martín0Steffen Lorenz Drees1Susanne Fetzner2Institute of Molecular Microbiology and Biotechnology, University of Münster, Corrensstraße 3, 48149 Münster, GermanyInstitute of Molecular Microbiology and Biotechnology, University of Münster, Corrensstraße 3, 48149 Münster, GermanyInstitute of Molecular Microbiology and Biotechnology, University of Münster, Corrensstraße 3, 48149 Münster, GermanyThe opportunistic pathogen <i>Pseudomonas aeruginosa</i> uses quorum sensing to control its virulence. One of its major signal molecules, the <i>Pseudomonas</i> quinolone signal PQS, has high affinity to membranes and is known to be trafficked mainly via outer membrane vesicles (OMVs). We previously reported that several 3-hydroxy-4(1<i>H</i>)-quinolone 2,4-dioxygenases (HQDs) catalyze the cleavage of PQS and thus act as quorum quenching enzymes. Further analysis showed that, in contrast to other HQDs, the activity of HQD from <i>Streptomyces bingchenggensis</i> (HQD<i><sub>S</sub>.<sub>b</sub>.</i>) was unexpectedly stabilized by culture supernatants of <i>P. aeruginosa</i>. Interestingly, the stabilizing effect was higher with supernatants from the strain PA14 than with supernatants from the strain PAO1. Heat treatment and lyophilization hardly affected the stabilizing effect; however, fractionation of the supernatant excluded small molecules as stabilizing agents. In a pull-down assay, HQD<i><sub>S</sub>.<sub>b</sub>.</i> appeared to interact with several <i>P. aeruginosa</i> proteins previously found in the OMV proteome. This prompted us to probe the physical interaction of HQD<i><sub>S</sub>.<sub>b</sub>.</i> with prepared extracellular membrane vesicles. Homo-FRET of fluorescently labeled HQD<i><sub>S</sub>.<sub>b</sub>.</i> indeed indicated a spatial clustering of the protein on the vesicles. Binding of a PQS-cleaving enzyme to the OMVs of <i>P. aeruginosa</i> may enhance PQS degradation and is highly reconcilable with its function as a quorum quenching enzyme.https://www.mdpi.com/2218-273X/12/11/1656dioxygenase<i>Pseudomonas</i> quinolone signal<i>Pseudomonas aeruginosa</i>quorum quenching enzymeouter membrane vesiclesmembrane-protein interaction |
| spellingShingle | Alba Arranz San Martín Steffen Lorenz Drees Susanne Fetzner A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i> Biomolecules dioxygenase <i>Pseudomonas</i> quinolone signal <i>Pseudomonas aeruginosa</i> quorum quenching enzyme outer membrane vesicles membrane-protein interaction |
| title | A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i> |
| title_full | A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i> |
| title_fullStr | A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i> |
| title_full_unstemmed | A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i> |
| title_short | A PQS-Cleaving Quorum Quenching Enzyme Targets Extracellular Membrane Vesicles of <i>Pseudomonas aeruginosa</i> |
| title_sort | pqs cleaving quorum quenching enzyme targets extracellular membrane vesicles of i pseudomonas aeruginosa i |
| topic | dioxygenase <i>Pseudomonas</i> quinolone signal <i>Pseudomonas aeruginosa</i> quorum quenching enzyme outer membrane vesicles membrane-protein interaction |
| url | https://www.mdpi.com/2218-273X/12/11/1656 |
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