Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests
ATP-binding cassette (ABC) transporters, particularly those of subfamily B, are involved in cell detoxification, multidrug resistance, drug treatment pharmacodynamics, and also ecological adaptation. In this regard, ABCB transporters may play a decisive role in the co-evolution between plants and he...
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| Format: | Article |
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MDPI AG
2024-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/29/22/5272 |
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| _version_ | 1846152845834846208 |
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| author | Mario Röpcke Sha Lu Cäcilia Plate Fee Meinzer Antonia Lisiecki Susanne Dobler |
| author_facet | Mario Röpcke Sha Lu Cäcilia Plate Fee Meinzer Antonia Lisiecki Susanne Dobler |
| author_sort | Mario Röpcke |
| collection | DOAJ |
| description | ATP-binding cassette (ABC) transporters, particularly those of subfamily B, are involved in cell detoxification, multidrug resistance, drug treatment pharmacodynamics, and also ecological adaptation. In this regard, ABCB transporters may play a decisive role in the co-evolution between plants and herbivores. Cardenolides, toxic steroid glycosides, are secondary plant metabolites that defend plants against herbivores by targeting their sodium–potassium ATPase. Despite their toxicity, several herbivorous insects such as the large milkweed bug (<i>Oncopeltus fasciatus</i>) have evolved adaptations to tolerate cardenolides and sequester them for their own defense. We investigate the role of two ABCB transporters of <i>O. fasciatus</i> for the paracellular transport of cardenolides by docking simulations and ATPase assays. Cardenolide binding of <i>Of</i>ABCB1 and <i>Of</i>ABCB2 is predicted by docking simulations and calculated binding energies are compared with substrate specificities determined in ATPase assays. Both tested ABCB transporters showed activity upon exposure to cardenolides and Km values that agreed well with the predictions of our docking simulations. We conclude that docking simulations can help identify transporter binding regions and predict substrate specificity, as well as provide deeper insights into the structural basis of ABC transporter function. |
| format | Article |
| id | doaj-art-4ff8e9b1e3e84b0b9d6361db5df946e3 |
| institution | Kabale University |
| issn | 1420-3049 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-4ff8e9b1e3e84b0b9d6361db5df946e32024-11-26T18:15:22ZengMDPI AGMolecules1420-30492024-11-012922527210.3390/molecules29225272Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental TestsMario Röpcke0Sha Lu1Cäcilia Plate2Fee Meinzer3Antonia Lisiecki4Susanne Dobler5Institute of Cell and System Biology of Animals, Universität Hamburg, 20146 Hamburg, GermanyInstitute of Cell and System Biology of Animals, Universität Hamburg, 20146 Hamburg, GermanyInstitute of Cell and System Biology of Animals, Universität Hamburg, 20146 Hamburg, GermanyInstitute of Cell and System Biology of Animals, Universität Hamburg, 20146 Hamburg, GermanyInstitute of Cell and System Biology of Animals, Universität Hamburg, 20146 Hamburg, GermanyInstitute of Cell and System Biology of Animals, Universität Hamburg, 20146 Hamburg, GermanyATP-binding cassette (ABC) transporters, particularly those of subfamily B, are involved in cell detoxification, multidrug resistance, drug treatment pharmacodynamics, and also ecological adaptation. In this regard, ABCB transporters may play a decisive role in the co-evolution between plants and herbivores. Cardenolides, toxic steroid glycosides, are secondary plant metabolites that defend plants against herbivores by targeting their sodium–potassium ATPase. Despite their toxicity, several herbivorous insects such as the large milkweed bug (<i>Oncopeltus fasciatus</i>) have evolved adaptations to tolerate cardenolides and sequester them for their own defense. We investigate the role of two ABCB transporters of <i>O. fasciatus</i> for the paracellular transport of cardenolides by docking simulations and ATPase assays. Cardenolide binding of <i>Of</i>ABCB1 and <i>Of</i>ABCB2 is predicted by docking simulations and calculated binding energies are compared with substrate specificities determined in ATPase assays. Both tested ABCB transporters showed activity upon exposure to cardenolides and Km values that agreed well with the predictions of our docking simulations. We conclude that docking simulations can help identify transporter binding regions and predict substrate specificity, as well as provide deeper insights into the structural basis of ABC transporter function.https://www.mdpi.com/1420-3049/29/22/5272ATP-binding cassette (ABC) transporterlarge milkweed bug<i>Oncopeltus fassciatus</i>cardenolidesco-evolutionary relationshipsATPase activity assays |
| spellingShingle | Mario Röpcke Sha Lu Cäcilia Plate Fee Meinzer Antonia Lisiecki Susanne Dobler Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests Molecules ATP-binding cassette (ABC) transporter large milkweed bug <i>Oncopeltus fassciatus</i> cardenolides co-evolutionary relationships ATPase activity assays |
| title | Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests |
| title_full | Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests |
| title_fullStr | Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests |
| title_full_unstemmed | Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests |
| title_short | Substrate Specificity of ABCB Transporters Predicted by Docking Simulations Can Be Confirmed by Experimental Tests |
| title_sort | substrate specificity of abcb transporters predicted by docking simulations can be confirmed by experimental tests |
| topic | ATP-binding cassette (ABC) transporter large milkweed bug <i>Oncopeltus fassciatus</i> cardenolides co-evolutionary relationships ATPase activity assays |
| url | https://www.mdpi.com/1420-3049/29/22/5272 |
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