Crystallization and preliminary X-ray diffraction studies of the ISC-like [2Fe-2S] ferredoxin (FdxB) from Pseudomonas putida JCM 20004.

The iron-sulfur (Fe-S) cluster-biosynthesis (ISC) system of the gamma-proteobacterium Pseudomonas putida JCM 20004 contains a constitutively expressed vertebrate-type [2Fe-2S] ferredoxin, FdxB, which lacks the conserved free cysteine residue near the Fe-S cluster site that has been proposed to function in the catalysis of biological Fe-S cluster assembly in other bacterial homologues. Recombinant FdxB was heterologously overproduced in Escherichia coli, purified and crystallized in its oxidized form by the hanging-drop vapour-diffusion and streak-seeding methods using 1.6 M trisodium citrate dihydrate pH 6.5. The thin needle-shaped crystals diffract to 1.90 A resolution and belong to the hexagonal space group P6(1)22, with unit-cell parameters a = 87.58, c = 73.14 A. The asymmetric unit contains one protein molecule.

[1]  D. Ohmori.  Characterization and reconstitution of Pseudomonas ovalis ferredoxin , 1984 .

[2]  A. Vagin,et al.  MOLREP: an Automated Program for Molecular Replacement , 1997 .

[3]  V. Stojanoff,et al.  Structure of a [2Fe–2S] ferredoxin from Rhodobacter capsulatus likely involved in Fe–S cluster biogenesis and conformational changes observed upon reduction , 2006, JBIC Journal of Biological Inorganic Chemistry.

[4]  R. Lill,et al.  Iron-sulfur-protein biogenesis in eukaryotes. , 2005, Trends in biochemical sciences.

[5]  H. Matsubara,et al.  Network of protein-protein interactions among iron-sulfur cluster assembly proteins in Escherichia coli. , 2002, Journal of biochemistry.

[6]  M. H. Barros,et al.  YAH1 of Saccharomyces cerevisiae: a new essential gene that codes for a protein homologous to human adrenodoxin. , 1999, Gene.

[7]  B. Matthews.  Solvent content of protein crystals. , 1968, Journal of molecular biology.

[8]  Y. Takahashi,et al.  Crystal structure of Escherichia coli Fdx, an adrenodoxin-type ferredoxin involved in the assembly of iron-sulfur clusters. , 2001, Biochemistry.

[9]  N. Cosper,et al.  Engineering a Three-cysteine, One-histidine Ligand Environment into a New Hyperthermophilic Archaeal Rieske-type [2Fe-2S] Ferredoxin from Sulfolobus solfataricus* , 2004, Journal of Biological Chemistry.

[10]  D. Dean,et al.  Structure, function, and formation of biological iron-sulfur clusters. , 2005, Annual review of biochemistry.

[11]  A. Crofts,et al.  Characterization of the pH-dependent resonance Raman transitions of archaeal and bacterial Rieske [2Fe-2S] proteins. , 2004, Journal of the American Chemical Society.

[12]  H. Beinert,et al.  Iron-sulfur clusters: nature's modular, multipurpose structures. , 1997, Science.