Structural Studies of Self-Assembled Subviral Particles: Combining Cell-Free Expression with 110 kHz MAS NMR Spectroscopy.

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.

• Authors: Guillaume David, Marie-Laure Fogeron, Maarten Schledorn, Roland Montserret, Uta Haselmann, Susanne Penzel, Aurélie Badillo, Lauriane Lecoq, Patrice André, Michael Nassal, Ralf Bartenschlager, Beat H Meier, Anja Böckmann
• Journal: Angewandte Chemie (International ed. in English) (Angew Chem Int Ed Engl) Vol. 57 Issue 17 Pg. 4787-4791 (04 16 2018) ISSN: 1521-3773 [Electronic] Germany
• PMID: 29457857 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chemical References

• Viral Matrix Proteins

Topics

• Cell-Free System
• Hepatitis B virus (chemistry)
• Nuclear Magnetic Resonance, Biomolecular
• Protein Conformation
• Viral Matrix Proteins (chemistry)