Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin
The Bordetella pertussis toxin (PT) is one important virulence factor causing the severe childhood disease whooping cough which still accounted for approximately 63,000 deaths worldwide in children in 2013. PT consists of PTS1, the enzymatically active (A) subunit and a non-covalently linked pentame...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2018-05-01
|
| Series: | Toxins |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2072-6651/10/5/181 |
| _version_ | 1811187131517763584 |
|---|---|
| author | Katharina Ernst Nina Eberhardt Ann-Katrin Mittler Michael Sonnabend Anna Anastasia Simon Freisinger Cordelia Schiene-Fischer Miroslav Malešević Holger Barth |
| author_facet | Katharina Ernst Nina Eberhardt Ann-Katrin Mittler Michael Sonnabend Anna Anastasia Simon Freisinger Cordelia Schiene-Fischer Miroslav Malešević Holger Barth |
| author_sort | Katharina Ernst |
| collection | DOAJ |
| description | The Bordetella pertussis toxin (PT) is one important virulence factor causing the severe childhood disease whooping cough which still accounted for approximately 63,000 deaths worldwide in children in 2013. PT consists of PTS1, the enzymatically active (A) subunit and a non-covalently linked pentameric binding/transport (B) subunit. After endocytosis, PT takes a retrograde route to the endoplasmic reticulum (ER), where PTS1 is released into the cytosol. In the cytosol, PTS1 ADP-ribosylates inhibitory alpha subunits of trimeric GTP-binding proteins (Giα) leading to increased cAMP levels and disturbed signalling. Here, we show that the cyclophilin (Cyp) isoforms CypA and Cyp40 directly interact with PTS1 in vitro and that Cyp inhibitors cyclosporine A (CsA) and its tailored non-immunosuppressive derivative VK112 both inhibit intoxication of CHO-K1 cells with PT, as analysed in a morphology-based assay. Moreover, in cells treated with PT in the presence of CsA, the amount of ADP-ribosylated Giα was significantly reduced and less PTS1 was detected in the cytosol compared to cells treated with PT only. The results suggest that the uptake of PTS1 into the cytosol requires Cyps. Therefore, CsA/VK112 represent promising candidates for novel therapeutic strategies acting on the toxin level to prevent the severe, life-threatening symptoms caused by PT. |
| first_indexed | 2024-04-11T13:57:01Z |
| format | Article |
| id | doaj.art-56e7cda1a3f14d4ead92af9aa3c6a609 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T13:57:01Z |
| publishDate | 2018-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-56e7cda1a3f14d4ead92af9aa3c6a6092022-12-22T04:20:13ZengMDPI AGToxins2072-66512018-05-0110518110.3390/toxins10050181toxins10050181Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis ToxinKatharina Ernst0Nina Eberhardt1Ann-Katrin Mittler2Michael Sonnabend3Anna Anastasia4Simon Freisinger5Cordelia Schiene-Fischer6Miroslav Malešević7Holger Barth8Institute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyInstitute for Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), GermanyInstitute of Pharmacology and Toxicology, University of Ulm Medical Center, 89081 Ulm, GermanyThe Bordetella pertussis toxin (PT) is one important virulence factor causing the severe childhood disease whooping cough which still accounted for approximately 63,000 deaths worldwide in children in 2013. PT consists of PTS1, the enzymatically active (A) subunit and a non-covalently linked pentameric binding/transport (B) subunit. After endocytosis, PT takes a retrograde route to the endoplasmic reticulum (ER), where PTS1 is released into the cytosol. In the cytosol, PTS1 ADP-ribosylates inhibitory alpha subunits of trimeric GTP-binding proteins (Giα) leading to increased cAMP levels and disturbed signalling. Here, we show that the cyclophilin (Cyp) isoforms CypA and Cyp40 directly interact with PTS1 in vitro and that Cyp inhibitors cyclosporine A (CsA) and its tailored non-immunosuppressive derivative VK112 both inhibit intoxication of CHO-K1 cells with PT, as analysed in a morphology-based assay. Moreover, in cells treated with PT in the presence of CsA, the amount of ADP-ribosylated Giα was significantly reduced and less PTS1 was detected in the cytosol compared to cells treated with PT only. The results suggest that the uptake of PTS1 into the cytosol requires Cyps. Therefore, CsA/VK112 represent promising candidates for novel therapeutic strategies acting on the toxin level to prevent the severe, life-threatening symptoms caused by PT.http://www.mdpi.com/2072-6651/10/5/181pertussis toxinwhooping coughcellular uptakemembrane transportcyclosporine Acyclophilinschaperonesnovel drug targets |
| spellingShingle | Katharina Ernst Nina Eberhardt Ann-Katrin Mittler Michael Sonnabend Anna Anastasia Simon Freisinger Cordelia Schiene-Fischer Miroslav Malešević Holger Barth Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin Toxins pertussis toxin whooping cough cellular uptake membrane transport cyclosporine A cyclophilins chaperones novel drug targets |
| title | Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin |
| title_full | Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin |
| title_fullStr | Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin |
| title_full_unstemmed | Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin |
| title_short | Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin |
| title_sort | pharmacological cyclophilin inhibitors prevent intoxication of mammalian cells with bordetella pertussis toxin |
| topic | pertussis toxin whooping cough cellular uptake membrane transport cyclosporine A cyclophilins chaperones novel drug targets |
| url | http://www.mdpi.com/2072-6651/10/5/181 |
| work_keys_str_mv | AT katharinaernst pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT ninaeberhardt pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT annkatrinmittler pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT michaelsonnabend pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT annaanastasia pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT simonfreisinger pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT cordeliaschienefischer pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT miroslavmalesevic pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin AT holgerbarth pharmacologicalcyclophilininhibitorspreventintoxicationofmammaliancellswithbordetellapertussistoxin |