Brief Research Report ARTICLE
100 kHz MAS proton-detected NMR spectroscopy of hepatitis B virus capsids
- 1UMR5086 Microbiologie Moléculaire et Biochimie Structurale (MMSB), France
- 2ETH Zürich, Switzerland
- 3University of Freiburg, Germany
We sequentially assigned the fully-protonated capsids made from core proteins of the Hepatitis B virus using proton detection at 100 kHz magic-angle spinning (MAS) in 0.7 mm rotors and compare sensitivity and assignment completeness to previously obtained assignments using carbon-detection techniques in 3.2 mm rotors and 17.5 kHz MAS. We show that proton detection shows a global gain of a factor ~50 in mass sensitivity, but that signal-to-noise ratios and completeness of the assignment was somewhat higher for carbon-detected experiments for comparable experimental times. We also show that deuteration and HN back protonation improves the proton linewidth at 100 kHz MAS by a factor of 1.5, from an average of 170 to 110 Hz, and by a factor of 1.3 compared to deuterated capsids at 60 kHz MAS in a 1.3 mm rotor. Yet, several HN protons cannot be back-exchanged due to solvent inaccessibility, which results in a total of 15 % of the amides missing in the spectra.
Keywords: solid-state NMR, Fast MAS, Proton detection, Carbon detection, Deuteration, Hepatitis B virus, Capsid, Core Protein
Received: 11 Apr 2019;
Accepted: 08 Jul 2019.
Edited by:Qian Han, Hainan University, China
Reviewed by:Kristaps Jaudzems, Latvian Institute of Organic Synthesis (LAS), Latvia
Loren B. Andreas, Max Planck Institute for Biophysical Chemistry, Germany
Copyright: © 2019 Lecoq, Schledorn, Wang, Smith-Penzel, Malär, Callon, Nassal, Meier and Böckmann. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Prof. Beat H. Meier, ETH Zürich, Zurich, 8092, Zürich, Switzerland, email@example.com
Dr. Anja Böckmann, UMR5086 Microbiologie Moléculaire et Biochimie Structurale (MMSB), Lyon, Rhône-Alpes, France, firstname.lastname@example.org