Expression, purification, crystallization and preliminary X-ray crystallographic studies of hepatitis B virus core fusion protein corresponding to octahedral particles

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Feb 1;69(Pt 2):165-9. doi: 10.1107/S1744309112052074. Epub 2013 Jan 31.

Abstract

Recombinant hepatitis B virus core proteins dimerize to form building blocks that are capable of self-assembly into a capsid. A core capsid protein dimer (CPD) linked to a green fluorescent protein variant, EGFP, at the C-terminus has been designed. The recombinant fusion CPD was expressed in Escherichia coli, assembled into virus-like particles (VLPs), purified and crystallized. The single crystal diffracted to 2.15 Å resolution and belonged to the cubic space group F432, with unit-cell parameters a = b = c = 219.7 Å. The fusion proteins assembled into icosahedral VLPs in aqueous solution, but were rearranged into octahedral symmetry through the crystal-packing process under the crystallization conditions.

Keywords: Escherichia coli; fusion protein; green fluorescent protein; hepatitis B virus; octahedral symmetry; virus-like particle.

MeSH terms

  • Capsid / chemistry
  • Crystallization
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Hepatitis B virus / metabolism*
  • Hepatitis B virus / ultrastructure
  • Microscopy, Fluorescence
  • Protein Multimerization
  • Ultracentrifugation
  • Viral Fusion Proteins / chemistry*
  • Viral Fusion Proteins / isolation & purification*
  • Virion / ultrastructure

Substances

  • Viral Fusion Proteins