دورية أكاديمية
Cell-free expression, purification, and membrane reconstitution for NMR studies of the nonstructural protein 4B from hepatitis C virus
| العنوان: | Cell-free expression, purification, and membrane reconstitution for NMR studies of the nonstructural protein 4B from hepatitis C virus |
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| المؤلفون: | Böckmann, Anja, Fogeron, Marie-Laure, Jirasko, Vlastimil, Penzel, Susanne, Paul, David, Montserret, Roland, Danis, Clément, Lacabanne, Denis, Badillo, Aurélie, Gouttenoire, Jérôme, Moradpour, Darius, Bartenschlager, Ralf, Penin, François, Meier, Beat H. |
| المصدر: | Journal of Biomolecular NMR, 65 (2) |
| بيانات النشر: | Springer |
| سنة النشر: | 2016 |
| المجموعة: | ETH Zürich Research Collection |
| مصطلحات موضوعية: | Cell-free protein expression, Integral membrane protein, Isotope labeling, Lipid reconstitution, NS4B, Solid-state NMR |
| الوصف: | We describe the expression of the hepatitis C virus nonstructural protein 4B (NS4B), which is an integral membrane protein, in a wheat germ cell-free system, the subsequent purification and characterization of NS4B and its insertion into proteoliposomes in amounts sufficient for multidimensional solid-state NMR spectroscopy. First spectra of the isotopically [2H,13C,15N]-labeled protein are shown to yield narrow 13C resonance lines and a proper, predominantly α-helical fold. Clean residue-selective leucine, isoleucine and threonine-labeling is demonstrated. These results evidence the suitability of the wheat germ-produced integral membrane protein NS4B for solid-state NMR. Still, the proton linewidth under fast magic angle spinning is broader than expected for a perfect sample and possible causes are discussed. ; ISSN:0925-2738 ; ISSN:1573-5001 |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/application/pdf |
| اللغة: | English |
| العلاقة: | info:eu-repo/grantAgreement/SNF/Projektförderung in Mathematik, Natur- und Ingenieurwissenschaften (Abteilung II)/146757; info:eu-repo/grantAgreement/SNF/Projekte MINT/159707; http://hdl.handle.net/20.500.11850/118305Test |
| DOI: | 10.3929/ethz-b-000118305 |
| الإتاحة: | https://doi.org/20.500.11850/118305Test https://doi.org/10.3929/ethz-b-000118305Test https://doi.org/10.1007/s10858-016-0040-2Test https://hdl.handle.net/20.500.11850/118305Test |
| حقوق: | info:eu-repo/semantics/openAccess ; http://rightsstatements.org/page/InC-NC/1.0Test/ ; In Copyright - Non-Commercial Use Permitted |
| رقم الانضمام: | edsbas.DA8F4F5C |
| قاعدة البيانات: | BASE |
| DOI: | 10.3929/ethz-b-000118305 |
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