Purification, crystallization and preliminary X-ray analysis of immunogenic virus-like particles formed by infectious bursal disease virus (IBDV) structural protein VP2

Acta Crystallogr D Biol Crystallogr. 2003 Jul;59(Pt 7):1234-7. doi: 10.1107/s090744490300859x. Epub 2003 Jun 27.

Abstract

Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry. VP2 protein, which consists of 452 amino-acid residues, is the primary immunogen of IBDV and contains the epitopes responsible for eliciting neutralizing antibodies. When the chimeric VP2 protein (rVP2H) of a local IBDV strain P3009 was expressed alone using the baculovirus system, virus-like particles of approximately 23 nm in diameter formed spontaneously. Highly pure rVP2H particles, obtained using ammonium sulfate precipitation, immobilized metal-ion affinity chromatography and gel-filtration chromatography, were successfully crystallized using the vapour-diffusion method. These crystals, with a maximum dimension of 0.4 mm, diffracted X-rays to 4.5 A resolution, but data were only collected to 6 A. Preliminary analysis of the diffraction data showed that the rVP2H crystals belong to the cubic space group P2(1)3, with unit-cell parameter 323.1 A. The icosahedral symmetry of the particles is clearly seen in the self-rotation function maps, with dyads and triads coincident with the crystallographic axes. Each asymmetric unit contains 1/3 of the particle, or 20 rVP2H subunits, and there are four particles in a unit cell, probably in a tetrahedral arrangement.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Baculoviridae / genetics
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Infectious bursal disease virus / chemistry*
  • Recombinant Proteins
  • Viral Structural Proteins / chemistry*
  • Viral Structural Proteins / immunology
  • Virion / chemistry*
  • Virion / immunology
  • Virion / isolation & purification

Substances

  • Recombinant Proteins
  • VP2 protein, infectious bursal disease virus
  • Viral Structural Proteins