Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier
The drug Rimantadine binds to two different sites in the M2 protein from influenza A, a peripheral site and a pore site that is the primary site of efficacy. It remained enigmatic that pore binding did not occur in certain detergent micelles, and in particular incomplete binding was observed in a mi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | Journal of Structural Biology: X |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590152423000065 |
| _version_ | 1797422499246374912 |
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| author | Kumar Tekwani Movellan Melanie Wegstroth Kerstin Overkamp Andrei Leonov Stefan Becker Loren B. Andreas |
| author_facet | Kumar Tekwani Movellan Melanie Wegstroth Kerstin Overkamp Andrei Leonov Stefan Becker Loren B. Andreas |
| author_sort | Kumar Tekwani Movellan |
| collection | DOAJ |
| description | The drug Rimantadine binds to two different sites in the M2 protein from influenza A, a peripheral site and a pore site that is the primary site of efficacy. It remained enigmatic that pore binding did not occur in certain detergent micelles, and in particular incomplete binding was observed in a mixture of lipids selected to match the viral membrane. Here we show that two effects are responsible, namely changes in the protein upon pore binding that prevented detergent solubilization, and slow binding kinetics in the lipid samples. Using 55–100 kHz magic-angle spinning NMR, we characterize kinetics of drug binding in three different lipid environments: DPhPC, DPhPC with cholesterol and viral mimetic membrane lipid bilayers. Slow pharmacological binding kinetics allowed the characterization of spectral changes associated with non-specific binding to the protein periphery in the kinetically trapped pore-apo state. Resonance assignments were determined from a set of proton-detected 3D spectra. Chemical shift changes associated with functional binding in the pore of M2 were tracked in real time in order to estimate the activation energy. The binding kinetics are affected by pH and the lipid environment and in particular cholesterol. We found that the imidazole-imidazole hydrogen bond at residue histidine 37 is a stable feature of the protein across several lipid compositions. Pore binding breaks the imidazole-imidazole hydrogen bond and limits solubilization in DHPC detergent. |
| first_indexed | 2024-03-09T07:34:14Z |
| format | Article |
| id | doaj.art-6126c5d4268f427484adf5d7d99bc435 |
| institution | Directory Open Access Journal |
| issn | 2590-1524 |
| language | English |
| last_indexed | 2024-03-09T07:34:14Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Structural Biology: X |
| spelling | doaj.art-6126c5d4268f427484adf5d7d99bc4352023-12-03T05:42:44ZengElsevierJournal of Structural Biology: X2590-15242023-12-018100090Real-time tracking of drug binding to influenza A M2 reveals a high energy barrierKumar Tekwani Movellan0Melanie Wegstroth1Kerstin Overkamp2Andrei Leonov3Stefan Becker4Loren B. Andreas5Department of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, Göttingen, GermanyDepartment of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, Göttingen, GermanyDepartment of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, Göttingen, GermanyDepartment of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, Göttingen, GermanyDepartment of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, Göttingen, GermanyCorresponding author.; Department of NMR Based Structural Biology, Max Planck Institute for Multidisciplinary Sciences, Am Fassberg 11, Göttingen, GermanyThe drug Rimantadine binds to two different sites in the M2 protein from influenza A, a peripheral site and a pore site that is the primary site of efficacy. It remained enigmatic that pore binding did not occur in certain detergent micelles, and in particular incomplete binding was observed in a mixture of lipids selected to match the viral membrane. Here we show that two effects are responsible, namely changes in the protein upon pore binding that prevented detergent solubilization, and slow binding kinetics in the lipid samples. Using 55–100 kHz magic-angle spinning NMR, we characterize kinetics of drug binding in three different lipid environments: DPhPC, DPhPC with cholesterol and viral mimetic membrane lipid bilayers. Slow pharmacological binding kinetics allowed the characterization of spectral changes associated with non-specific binding to the protein periphery in the kinetically trapped pore-apo state. Resonance assignments were determined from a set of proton-detected 3D spectra. Chemical shift changes associated with functional binding in the pore of M2 were tracked in real time in order to estimate the activation energy. The binding kinetics are affected by pH and the lipid environment and in particular cholesterol. We found that the imidazole-imidazole hydrogen bond at residue histidine 37 is a stable feature of the protein across several lipid compositions. Pore binding breaks the imidazole-imidazole hydrogen bond and limits solubilization in DHPC detergent.http://www.sciencedirect.com/science/article/pii/S2590152423000065Magic-angle spinningProton channelDrug bindingSolid-state NMRBinding kinetics |
| spellingShingle | Kumar Tekwani Movellan Melanie Wegstroth Kerstin Overkamp Andrei Leonov Stefan Becker Loren B. Andreas Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier Journal of Structural Biology: X Magic-angle spinning Proton channel Drug binding Solid-state NMR Binding kinetics |
| title | Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier |
| title_full | Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier |
| title_fullStr | Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier |
| title_full_unstemmed | Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier |
| title_short | Real-time tracking of drug binding to influenza A M2 reveals a high energy barrier |
| title_sort | real time tracking of drug binding to influenza a m2 reveals a high energy barrier |
| topic | Magic-angle spinning Proton channel Drug binding Solid-state NMR Binding kinetics |
| url | http://www.sciencedirect.com/science/article/pii/S2590152423000065 |
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