Structural Studies of Self‐Assembled Subviral Particles: Combining Cell‐Free Expression with 110 kHz MAS NMR Spectroscopy. Issue 17 (25th March 2018)

• Record Type: Journal Article
• Authors: David, Guillaume; Fogeron, Marie‐Laure; Schledorn, Maarten; Montserret, Roland; Haselmann, Uta; Penzel, Susanne; Badillo, Aurélie; Lecoq, Lauriane; André, Patrice; Nassal, Michael; Bartenschlager, Ralf; Meier, Beat H.; Böckmann, Anja
• Is Part Of: Angewandte Chemie international edition. Volume 57:Issue 17(2018)
• Page Start: 4787
• Page End: 4791
• Publication Date: 2018-03-25
• Subjects: cell-free protein expression -- duck hepatitis B virus -- proton detection -- solid-state NMR spectroscopy -- subviral particles; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201712091
• Languages: English
• ISSNs: 1433-7851

Abstract

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. Abstract : By combining cell‐free protein expression with 110 kHz magic‐angle‐spinning solid‐state NMR spectroscopy, spectra were recorded for less than 500 μg of in vitro self‐assembled subviral particles of the duck hepatitis B virus. This method should pave the way for investigations of large membrane protein assemblies whose structures have not been solved thus far.