التفاصيل البيبلوغرافية
| العنوان: |
Cell-free expression, purification, and membrane reconstitution for NMR studies of the nonstructural protein 4B from hepatitis C virus. |
| المؤلفون: |
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, Böckmann, Anja |
| المصدر: |
Journal of Biomolecular NMR; Jun2016, Vol. 65 Issue 2, p87-98, 12p |
| مستخلص: |
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 [H,C,N]-labeled protein are shown to yield narrow C 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. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
