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The structure of the all-cysteinyl-coordinated D14C variant of [4Fe–4S] ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus has been determined to 1.7 Å resolution from a crystal belonging to space group C2221 with two types of molecules, A and B, in the asymmetric unit. A and B molecules have different crystal packing and intramolecular disulfide bond conformation. The crystal packing reveals a β-sheet interaction between A molecules in adjacent asymmetric units, whereas B molecules are packed as monomers in a less rigid position next to the A–A extended β-sheet dimers. The A molecules contain an intramolecular disulfide bond in a double conformation with 60% occupancy left-handed and 40% occupancy right-handed spiral conformation, whereas B molecules have an intramolecular disulfide bond in a right-handed spiral conformation. The cluster in D14C [4Fe–4S] P. furiosus ferredoxin was chemically oxidized at pH 5.8 to [3Fe–4S]. For purification at pH 8.0, two forms of the protein are obtained. Mass spectrometric analysis shows that the two forms are the D14C [3Fe–4S] P. furiosus ferredoxin monomer and a disulfide-bonded dimer of D14C [3Fe–4S] P. furiosus ferredoxin. When oxidization and purification are carried out at pH 5.8, only the monomer is obtained. The crystal structure of D14C [3Fe–4S] P. furiosus ferredoxin monomer was determined to 2.8 Å resolution from a crystal belonging to space group P212121 with two molecules in the asymmetric unit. The molecules resemble molecule A of D14C [4Fe–4S] P. furiosus ferredoxin and electron density clearly shows the presence of a [3Fe–4S] cluster.
Original languageEnglish
JournalJournal of Biological Inorganic Chemistry
Publication date2011
Volume16
Issue5
Pages763-775
ISSN0949-8257
DOIs
StateE-pub ahead of print

Bibliographical note

An interactive 3D complement page in Proteopedia is available at http://proteopedia.org/wiki/index.php/Journal:JBIC:10

CitationsWeb of Science® Times Cited: 2

Keywords

  • Disulfide bond, Crystal structure, Dimer, Ferredoxin, Pyrococcus furiosus
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