Angewandte Chemie - International Edition
Angewandte Chemie - International Edition
5 years ago

Towards structural studies of self-assembled subviral particles: combining cell-free expression with 100 kHz MAS NMR

Michael Nassal, Patrice André, Aurelie Badillo, Marie-Laure Fogeron, Guillaume David, Maarten Schledorn, Uta Haselmann, Susanne Penzel, Beat H Meier, Ralf Bartenschlager, Anja Böckmann, Lauriane Lecoq, Roland Montserret
Viral membrane proteins are prime targets in the combat against infection. Still, their structure determination remains a challenge, both with respect to sample preparation, and the need for structural methods allowing analysis in a native-like lipid environment. Cell-free protein synthesis and solid-state NMR are promising approaches in this context, one with respect to its high potential of native expression of complex proteins, and the other for its ability to analyze membrane proteins in lipids. We here show that milligram amounts of the small envelope protein of the duck hepatitis B virus (DHBV) can be produced using cell-free expression, and that the protein self-assembles into subviral particles. 2D proton-detected NMR spectra recorded at 110 kHz magic angle spinning on < 500 μg protein show a number of isolated peaks with linewidths comparable to model membrane proteins, paving the way for structural studies this homologous protein to a potential drug target in HBV infection.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/anie.201712091

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