Crystallization and preliminary X-ray diffraction studies of the ISC-like [2Fe–2S] ferredoxin (FdxB) from Pseudomonas putida JCM 20004
- Department of Biochemistry and Molecular Biology, Nippon Medical School, Sendagi, Bunkyo-ku, Tokyo 113-8602 (Japan)
- Department of Chemistry, Juntendo University, Inba, Chiba 270-1695 (Japan)
- Japan Synchrotron Radiation Research Institute (SPring-8/JASRI), Sayo, Hyogo 679-5198 (Japan)
A vertebrate-type [2Fe–2S] ferredoxin (FdxB), which is probably involved in the iron–sulfur cluster-biosynthesis system of the γ-proteobacterium P. putida JCM 20004, has been crystallized in space group P6{sub 1}22. The needle-shaped crystals of recombinant FdxB diffract to 1.90 Å resolution using synchrotron radiation. The iron–sulfur (Fe–S) cluster-biosynthesis (ISC) system of the γ-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 Å resolution and belong to the hexagonal space group P6{sub 1}22, with unit-cell parameters a = 87.58, c = 73.14 Å. The asymmetric unit contains one protein molecule.
- OSTI ID:
- 22360444
- Journal Information:
- Acta Crystallographica. Section F, Journal Name: Acta Crystallographica. Section F Journal Issue: Pt 12 Vol. 63; ISSN ACSFCL; ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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