Protein preparation, crystallization and preliminary X-ray analysis of Polygonum cuspidatum bifunctional chalcone synthase/benzalacetone synthase

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Aug;69(Pt 8):871-5. doi: 10.1107/S1744309113017004. Epub 2013 Jul 27.

Abstract

The chalcone synthase (CHS) superfamily of type III polyketide synthases (PKSs) generate the backbones of a variety of plant secondary metabolites. An active bifunctional chalcone synthase/benzalacetone synthase (CHS/BAS) from Polygonum cuspidatum was overexpressed in Escherichia coli as a C-terminally polyhistidine-tagged fusion protein, purified to homogeneity and crystallized using polyethylene glycol 4000 as a precipitant. The production of well shaped crystals of the complex between PcPKS1 and benzalacetone was dependent on the presence of sorbitol and barium chloride as additives. The crystals belonged to the orthorhombic space group P2₁2₁2₁, with unit-cell parameters a = 80.23, b = 81.01, c = 122.89 Å, and diffracted X-rays to at least 2.0 Å resolution.

Keywords: Polygonum cuspidatum; chalcone synthase/benzalacetone synthase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / chemistry*
  • Acyltransferases / genetics
  • Amino Acid Sequence
  • Butanones / chemistry*
  • Crystallization
  • Fallopia japonica / enzymology*
  • Fallopia japonica / genetics
  • Molecular Sequence Data
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • X-Ray Diffraction

Substances

  • Butanones
  • Plant Proteins
  • benzylideneacetone
  • Acyltransferases
  • flavanone synthetase