<i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages
The protein toxin C3bot from <i>Clostridium botulinum</i> is a mono-ADP-ribosyltransferase that selectively intoxicates monocyte-derived cells such as macrophages, osteoclasts, and dendritic cells (DCs) by cytosolic modification of Rho-A, -B, and -C. Here, we investigated the application...
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MDPI AG
2022-10-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/14/10/711 |
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| author | Maximilian Fellermann Mia Stemmer Reiner Noschka Fanny Wondany Stephan Fischer Jens Michaelis Steffen Stenger Holger Barth |
| author_facet | Maximilian Fellermann Mia Stemmer Reiner Noschka Fanny Wondany Stephan Fischer Jens Michaelis Steffen Stenger Holger Barth |
| author_sort | Maximilian Fellermann |
| collection | DOAJ |
| description | The protein toxin C3bot from <i>Clostridium botulinum</i> is a mono-ADP-ribosyltransferase that selectively intoxicates monocyte-derived cells such as macrophages, osteoclasts, and dendritic cells (DCs) by cytosolic modification of Rho-A, -B, and -C. Here, we investigated the application of C3bot as well as its non-toxic variant C3bot<sub>E174Q</sub> as transporters for selective delivery of cargo molecules into macrophages and DCs. C3bot and C3bot<sub>E174Q</sub> facilitated the uptake of eGFP into early endosomes of human-monocyte-derived macrophages, as revealed by stimulated emission depletion (STED) super-resolution microscopy. The fusion of the cargo model peptide eGFP neither affected the cell-type selectivity (enhanced uptake into human macrophages ex vivo compared to lymphocytes) nor the cytosolic release of C3bot. Moreover, by cell fractionation, we demonstrated that C3bot and C3bot<sub>E174Q</sub> strongly enhanced the cytosolic release of functional eGFP. Subsequently, a modular system was created on the basis of C3bot<sub>E174Q</sub> for covalent linkage of cargos via thiol–maleimide click chemistry. The functionality of this system was proven by loading small molecule fluorophores or an established reporter enzyme and investigating the cellular uptake and cytosolic release of cargo. Taken together, non-toxic C3bot<sub>E174Q</sub> is a promising candidate for the cell-type-selective delivery of small molecules, peptides, and proteins into the cytosol of macrophages and DCs. |
| first_indexed | 2024-03-09T19:24:40Z |
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| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-09T19:24:40Z |
| publishDate | 2022-10-01 |
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| record_format | Article |
| series | Toxins |
| spelling | doaj.art-2b66a39aac1e4a8d92a5e296924ca4d82023-11-24T02:59:33ZengMDPI AGToxins2072-66512022-10-01141071110.3390/toxins14100711<i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and MacrophagesMaximilian Fellermann0Mia Stemmer1Reiner Noschka2Fanny Wondany3Stephan Fischer4Jens Michaelis5Steffen Stenger6Holger Barth7Institute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute for Medical Microbiology and Hygiene, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Biophysics, Ulm University, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Biophysics, Ulm University, 89081 Ulm, GermanyInstitute for Medical Microbiology and Hygiene, University of Ulm Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, University of Ulm Medical Center, 89081 Ulm, GermanyThe protein toxin C3bot from <i>Clostridium botulinum</i> is a mono-ADP-ribosyltransferase that selectively intoxicates monocyte-derived cells such as macrophages, osteoclasts, and dendritic cells (DCs) by cytosolic modification of Rho-A, -B, and -C. Here, we investigated the application of C3bot as well as its non-toxic variant C3bot<sub>E174Q</sub> as transporters for selective delivery of cargo molecules into macrophages and DCs. C3bot and C3bot<sub>E174Q</sub> facilitated the uptake of eGFP into early endosomes of human-monocyte-derived macrophages, as revealed by stimulated emission depletion (STED) super-resolution microscopy. The fusion of the cargo model peptide eGFP neither affected the cell-type selectivity (enhanced uptake into human macrophages ex vivo compared to lymphocytes) nor the cytosolic release of C3bot. Moreover, by cell fractionation, we demonstrated that C3bot and C3bot<sub>E174Q</sub> strongly enhanced the cytosolic release of functional eGFP. Subsequently, a modular system was created on the basis of C3bot<sub>E174Q</sub> for covalent linkage of cargos via thiol–maleimide click chemistry. The functionality of this system was proven by loading small molecule fluorophores or an established reporter enzyme and investigating the cellular uptake and cytosolic release of cargo. Taken together, non-toxic C3bot<sub>E174Q</sub> is a promising candidate for the cell-type-selective delivery of small molecules, peptides, and proteins into the cytosol of macrophages and DCs.https://www.mdpi.com/2072-6651/14/10/711clostridial C3 toxin<i>Clostridium botulinum</i>C3botC3bot<sub>E174Q</sub>dendritic cellsmacrophages |
| spellingShingle | Maximilian Fellermann Mia Stemmer Reiner Noschka Fanny Wondany Stephan Fischer Jens Michaelis Steffen Stenger Holger Barth <i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages Toxins clostridial C3 toxin <i>Clostridium botulinum</i> C3bot C3bot<sub>E174Q</sub> dendritic cells macrophages |
| title | <i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages |
| title_full | <i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages |
| title_fullStr | <i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages |
| title_full_unstemmed | <i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages |
| title_short | <i>Clostridium botulinum</i> C3 Toxin for Selective Delivery of Cargo into Dendritic Cells and Macrophages |
| title_sort | i clostridium botulinum i c3 toxin for selective delivery of cargo into dendritic cells and macrophages |
| topic | clostridial C3 toxin <i>Clostridium botulinum</i> C3bot C3bot<sub>E174Q</sub> dendritic cells macrophages |
| url | https://www.mdpi.com/2072-6651/14/10/711 |
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