Solid-state [13C–15N] NMR resonance assignment of hepatitis B virus core protein
- 142 Downloads
Each year, nearly 900,000 deaths are due to serious liver diseases caused by chronic hepatitis B virus infection. The viral particle is composed of an outer envelope and an inner icosahedral nucleocapsid formed by multiple dimers of a ~ 20 kDa self-assembling core protein (Cp). Here we report the solid-state 13C and 15N resonance assignments of the assembly domain, Cp149, of the core protein in its capsid form. A secondary chemical shift analysis of the 140 visible residues suggests an overall alpha-helical three-dimensional fold matching that derived for Cp149 from the X-ray crystallography of the capsid, and from solution-state NMR of the Cp149 dimer. Interestingly, however, at three distinct regions the chemical shifts in solution differ significantly between core proteins in the capsid state versus in the dimer state, strongly suggesting the respective residues to be involved in capsid assembly.
KeywordsHepatitis B virus Core protein Nucleocapsid Solid-state NMR Assignments
This work was supported by the French ANR (ANR-14-CE09-0024B), the LABEX ECOFECT (ANR-11-LABX-0048) within the Université de Lyon program Investissements d’Avenir (ANR-11-IDEX-0007) (AB), the Swiss National Science Foundation (Grant 200020_159707) (BHM), the DFG Grant NA154/9-4 (MN), a doctoral stipend from the Chinese Scientific Council (SW) and by the Marie Skłodowska-Curie program (H2020-MSCA-IF-2016 748516) (LL). We thank the Centre d’Imagerie Quantitative Lyon-Est (CIQLE) for the access and training on the electron microscopy platform.
- Birnbaum F, Nassal M (1990) Hepatitis B virus nucleocapsid assembly: primary structure requirements in the core protein. J Virol 64:3319–3330Google Scholar
- Gallina A, Bonelli F, Zentilin L et al (1989) A recombinant hepatitis B core antigen polypeptide with the protamine-like domain deleted self-assembles into capsid particles but fails to bind nucleic acids. J Virol 63:4645–4652Google Scholar
- Heger-Stevic J, Kolb P, Walker A, Nassal M Displaying whole-chain proteins on hepatitis B virus capsid-like particles. In: Wege C, Lomonossoff G (eds) Virus-Derived Nanoparticles for Advanced Technologies, Chap 33. Springer, Berlin (in press)Google Scholar
- Nassal M (1992) The arginine-rich domain of the hepatitis B virus core protein is required for pregenome encapsidation and productive viral positive-strand DNA synthesis but not for virus assembly. J Virol 66:4107–4116Google Scholar
- Penzel S, Smith AA, Agarwal V et al (2015) Protein resonance assignment at MAS frequencies approaching 100 kHz: a quantitative comparison of J-coupling and dipolar- coupling-based transfer methods. J Biomol NMR 1–22. https://doi.org/10.1007/s10858-015-9975-y
- World Health Organization (2017) Global hepatitis report. http://www.who.int