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

Lauriane Lecoq (1) , Maarten Schledorn , Shishan Wang (2) , Susanne Smith-Penzel , Alexander A Malär , Morgane Callon (1) , Michael Nassal (3) , Beat Meier (4) , Anja Böckmann (2)

1 IBS - UMR 5075 - Institut de biologie structurale
2 MMSB - Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry
3 University Hospital Freiburg
4 ETH Zürich - Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich]

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 H N 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 H N protons cannot be back-exchanged due to solvent inaccessibility, which results in a total of 15% of the amides missing in the spectra.

Mots clés: solid-state NMR; fast MAS; proton detection; carbon detection; deuteration; hepatitis B virus; capsid; core protein

Domaines: Biologie structurale [q-bio.BM]; Chimie analytique

Lauriane Lecoq, Maarten Schledorn, Shishan Wang, Susanne Smith-Penzel, Alexander A Malär, et al.. 100 kHz MAS Proton-Detected NMR Spectroscopy of Hepatitis B Virus Capsids. Frontiers in Molecular Biosciences, 2019, 6, ⟨10.3389/fmolb.2019.00058⟩. ⟨hal-02322643⟩