Overexpression, purification, crystallization and preliminary X-ray diffraction analysis of Cu,Zn superoxide dismutase from Peking duck.
The cDNA encoding Peking duck Cu,Zn superoxide dismutase (dSOD) was cloned and sequenced. The recombinant enzyme was overexpressed in Escherichia coli, purified to homogeneity and crystallized using the sitting-drop vapour-diffusion technique. Trigonal crystals of dSOD were obtained at 278 K at low ionic strength and around neutral pH. These crystals belong to space group P3(2)21, with unit-cell parameters a = 124.4, c = 163.5 A, gamma = 120 degrees. The asymmetric unit contains four dimers (eight monomers of Cu,Zn dSOD) and has a 56% solvent content, with a V(M) of 2.8 A(3) Da(-1). On a Rigaku R-AXIS IIc image-plate area-detector system, the crystal diffracted to 2.9 A. Unusual supermolecular double-helix packing with 9(2)2 non-crystallographic symmetry in crystals has been observed in the initial structural analysis.
Liu
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W
Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
Li
P W
PW
Li
G P
GP
Zhu
R H
RH
Wang
D C
DC
D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D004372:Ducks; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification
D011994:Recombinant Proteins; D013482:Superoxide Dismutase