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100 kHz MAS Proton-Detected NMR Spectroscopy of Hepatitis B Virus Capsids.

  • Lecoq L Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon, Lyon, France.
  • Schledorn M Physical Chemistry, ETH Zurich, Zurich, Switzerland.
  • Wang S Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon, Lyon, France.
  • Smith-Penzel S Physical Chemistry, ETH Zurich, Zurich, Switzerland.
  • Malär AA Physical Chemistry, ETH Zurich, Zurich, Switzerland.
  • Callon M Physical Chemistry, ETH Zurich, Zurich, Switzerland.
  • Nassal M Department of Medicine II/Molecular Biology, Medical Center, University Hospital Freiburg, University of Freiburg, Freiburg, Germany.
  • Meier BH Physical Chemistry, ETH Zurich, Zurich, Switzerland.
  • Böckmann A Molecular Microbiology and Structural Biochemistry, Labex Ecofect, UMR 5086 CNRS, Université de Lyon, Lyon, France.
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  • 2019-08-10
Published in:
  • Frontiers in molecular biosciences. - 2019
capsid
carbon detection
core protein
deuteration
fast MAS
hepatitis B virus
proton detection
solid-state NMR
English 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 170-110 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.
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  • English
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https://sonar.ch/global/documents/238599
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