Crystallization and Preliminary X-ray Diffraction Studies of the ISC-like [2Fe-2S] Ferredoxin (FdxB) from Pseudomonas Putida JCM 20004

Overview

Journal Acta Crystallogr Sect F Struct Biol Cryst Commun

Date 2007 Dec 18

PMID 18084081

Citations 1

Authors

Toshio Iwasaki

Daijiro Ohmori

Nobutaka Shimizu

Takashi Kumasaka

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

ISC-like [2Fe-2S] ferredoxin (FdxB) dimer from Pseudomonas putida JCM 20004: structural and electron-nuclear double resonance characterization.

Iwasaki T, Kappl R, Bracic G, Shimizu N, Ohmori D, Kumasaka T J Biol Inorg Chem. 2011; 16(6):923-35.

PMID: 21647778 DOI: 10.1007/s00775-011-0793-8.

References

Nelson K, Weinel C, Paulsen I, Dodson R, HILBERT H, Martins Dos Santos V . Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. Environ Microbiol. 2003; 4(12):799-808. DOI: 10.1046/j.1462-2920.2002.00366.x. View

BEINERT H . Iron-sulfur proteins: ancient structures, still full of surprises. J Biol Inorg Chem. 2000; 5(1):2-15. DOI: 10.1007/s007750050002. View

Kounosu A, Li Z, Cosper N, Shokes J, Scott R, Imai T . Engineering a three-cysteine, one-histidine ligand environment into a new hyperthermophilic archaeal Rieske-type [2Fe-2S] ferredoxin from Sulfolobus solfataricus. J Biol Chem. 2004; 279(13):12519-28. DOI: 10.1074/jbc.M305923200. View

Ohmori D, Yamakura F, Suzuki K . Purification and some properties of a 2Fe ferredoxin in Pseudomonas ovalis. Biochem Int. 1989; 18(3):573-80. View

Johnson D, Dean D, Smith A, Johnson M . Structure, function, and formation of biological iron-sulfur clusters. Annu Rev Biochem. 2005; 74:247-81. DOI: 10.1146/annurev.biochem.74.082803.133518. View

Rouault T, Tong W . Iron-sulphur cluster biogenesis and mitochondrial iron homeostasis. Nat Rev Mol Cell Biol. 2005; 6(4):345-51. DOI: 10.1038/nrm1620. View

BEINERT H, Holm R, Munck E . Iron-sulfur clusters: nature's modular, multipurpose structures. Science. 1997; 277(5326):653-9. DOI: 10.1126/science.277.5326.653. View

Tokumoto U, Nomura S, Minami Y, Mihara H, Kato S, Kurihara T . Network of protein-protein interactions among iron-sulfur cluster assembly proteins in Escherichia coli. J Biochem. 2002; 131(5):713-9. DOI: 10.1093/oxfordjournals.jbchem.a003156. View

Matthews B . Solvent content of protein crystals. J Mol Biol. 1968; 33(2):491-7. DOI: 10.1016/0022-2836(68)90205-2. View

10.

Sainz G, Jakoncic J, Sieker L, Stojanoff V, Sanishvili N, Asso M . Structure of a [2Fe-2S] ferredoxin from Rhodobacter capsulatus likely involved in Fe-S cluster biogenesis and conformational changes observed upon reduction. J Biol Inorg Chem. 2006; 11(2):235-46. DOI: 10.1007/s00775-005-0069-2. View

11.

Lill R, Muhlenhoff U . Iron-sulfur-protein biogenesis in eukaryotes. Trends Biochem Sci. 2005; 30(3):133-41. DOI: 10.1016/j.tibs.2005.01.006. View

12.

Barros M, Nobrega F . YAH1 of Saccharomyces cerevisiae: a new essential gene that codes for a protein homologous to human adrenodoxin. Gene. 1999; 233(1-2):197-203. DOI: 10.1016/s0378-1119(99)00137-7. View

13.

Kakuta Y, Horio T, Takahashi Y, Fukuyama K . Crystal structure of Escherichia coli Fdx, an adrenodoxin-type ferredoxin involved in the assembly of iron-sulfur clusters. Biochemistry. 2001; 40(37):11007-12. DOI: 10.1021/bi010544t. View

14.

Iwasaki T, Kounosu A, Kolling D, Crofts A, Dikanov S, Jin A . Characterization of the pH-dependent resonance Raman transitions of archaeal and bacterial Rieske [2Fe-2S] proteins. J Am Chem Soc. 2004; 126(15):4788-9. DOI: 10.1021/ja031976p. View