Structural Studies of Self‐Assembled Subviral Particles: Combining Cell‐Free Expression with 110 kHz MAS NMR Spectroscopy |
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| Authors: | Guillaume David Dr Marie‐Laure Fogeron Maarten Schledorn Roland Montserret Uta Haselmann Susanne Penzel Dr Aurélie Badillo Dr Lauriane Lecoq Prof Patrice André Prof Michael Nassal Prof Ralf Bartenschlager Prof Beat H Meier Dr Anja Böckmann |
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| Institution: | 1. Institut de Biologie et Chimie des Protéines, MMSB, Labex Ecofect, UMR 5086 CNRS, Université de Lyon, Lyon, France;2. Physical Chemistry, ETH Zurich, Zurich, Switzerland;3. Department of Infectious Diseases, Molecular Virology, Heidelberg University, Heidelberg, Germany;4. Division of Virus-Associated Carcinogenesis Germany, Cancer Research Center (DKFZ), Heidelberg, Germany;5. RD-Biotech, Recombinant Protein Unit, Besan?on, France;6. Centre International de Recherche en Infectiologie, Institut National de la Santé et de la Recherche Médicale Unité 1111, Centre National de la Recherche Scientifique Unités Mixte de Recherche, Lyon, France;7. Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Villeurbanne, France;8. Université de Lyon, Laboratoire de Virologie, H?pital de la Croix-Rousse, Hospices Civils de Lyon, Lyon, France;9. University Hospital Freiburg, Internal Medicine II/Molecular Biology, Freiburg, Germany |
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| Abstract: | Viral membrane proteins are prime targets in combatting infection. Still, the determination of their structure remains a challenge, both with respect to sample preparation and the need for structural methods allowing for analysis in a native‐like lipid environment. Cell‐free protein synthesis and solid‐state NMR spectroscopy are promising approaches in this context, the former with respect to its great potential in the native expression of complex proteins, and the latter for the analysis of membrane proteins in lipids. Herein, we show that milligram amounts of the small envelope protein of the duck hepatitis B virus (DHBV) can be produced by cell‐free expression, and that the protein self‐assembles into subviral particles. Proton‐detected 2D NMR spectra recorded at a magic‐angle‐spinning frequency of 110 kHz on <500 μg protein show a number of isolated peaks with line widths comparable to those of model membrane proteins, paving the way for structural studies of this protein that is homologous to a potential drug target in HBV infection. |
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| Keywords: | cell-free protein expression duck hepatitis B virus proton detection solid-state NMR spectroscopy subviral particles |
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