Crystallization and preliminary X-ray diffraction studies of a hyperthermophilic Rieske protein variant (SDX-triple) with an engineered rubredoxin-like mononuclear iron site

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Oct 1;62(Pt 10):993-5. doi: 10.1107/S1744309106034476. Epub 2006 Sep 30.

Abstract

In place of the Rieske [2Fe-2S] cluster, an archetypal mononuclear iron site has rationally been designed into a hyperthermophilic archaeal Rieske [2Fe-2S] protein (sulredoxin) from Sulfolobus tokodaii by three residue replacements with reference to the Pyrococcus furiosus rubredoxin sequence. The resulting sulredoxin variant, SDX-triple (H44I/A45C/H64C), has been purified and crystallized by the hanging-drop vapour-diffusion method using 65%(v/v) 2-methyl-2,4-pentanediol, 0.025 M citric acid and 0.075 M sodium acetate trihydrate pH 4.3. The crystals diffract to 1.63 A resolution and belong to the triclinic space group P1, with unit-cell parameters a = 43.56, b = 76.54, c = 80.28 A, alpha = 88.12, beta = 78.82, gamma = 73.46 degrees. The asymmetric unit contains eight protein molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Crystallization
  • Crystallography, X-Ray
  • Electron Transport Complex III / chemistry*
  • Electron Transport Complex III / genetics
  • Iron / chemistry*
  • Iron / metabolism
  • Iron-Sulfur Proteins / chemistry*
  • Iron-Sulfur Proteins / genetics
  • Molecular Sequence Data
  • Protein Engineering
  • Pyrococcus furiosus / chemistry
  • Pyrococcus furiosus / enzymology
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Rubredoxins / chemistry*
  • Rubredoxins / genetics
  • Sequence Alignment
  • Sulfolobus / chemistry
  • Sulfolobus / enzymology
  • X-Ray Diffraction

Substances

  • Iron-Sulfur Proteins
  • Recombinant Proteins
  • Rieske iron-sulfur protein
  • Rubredoxins
  • Iron
  • Electron Transport Complex III