Crystal structures of the all-cysteinyl-coordinated D14C variant of Pyrococcus furiosus ferredoxin: [4Fe-4S] a†" [3Fe-4S] cluster conversion

被引:9
作者
Lovgreen, Monika Nohr [1 ]
Martic, Maja [1 ]
Windahl, Michael S. [2 ]
Christensen, Hans E. M. [1 ]
Harris, Pernille [1 ]
机构
[1] Tech Univ Denmark, Dept Chem, DK-2800 Lyngby, Denmark
[2] Carlsberg Lab, DK-1799 Copenhagen V, Denmark
来源
JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY | 2011年 / 16卷 / 05期
关键词
Ferredoxin; Crystal structure; Pyrococcus furiosus; Disulfide bond; Dimer; IRON-SULFUR CLUSTER; BACILLUS-THERMOPROTEOLYTICUS; SPECTROSCOPIC CHARACTERIZATION; DESULFOVIBRIO-AFRICANUS; RESOLUTION STRUCTURES; ELECTRON-TRANSFER; PROTEIN; ASPARTATE; REDUCTION; REDOX;
D O I
10.1007/s00775-011-0778-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 resolution from a crystal belonging to space group C222(1) 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 beta-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 beta-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 resolution from a crystal belonging to space group P2(1)2(1)2(1) 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.
引用
收藏
页码:763 / 775
页数:13
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