Crystallization and preliminary X‐ray analysis of the monomeric Cu, Zn superoxide dismutase from Escherichia coli

• Published in: Protein science 1996-10, Vol.5 (10), p.2125-2127
• Main Authors: Battistoni, Andrea, Folcarelli, Silvia, Rotilio, Giuseppe, Desideri, Alessandro, Capasso, Clemente, Pesce, Alessandra, Bolognesi, Martino
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Copper - chemistry; Crystallization; Crystallography, X-Ray; crystals; Dimerization; enzyme structure; Escherichia coli; Escherichia coli - enzymology; Fungal Proteins - chemistry; metal proteins; Molecular Sequence Data; monomer‐dimer equilibrium; Polyethylene Glycols; Protein Conformation; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - isolation & purification; Sequence Alignment; Sequence Homology, Amino Acid; Species Specificity; Superoxide Dismutase - chemistry; Superoxide Dismutase - isolation & purification; Vertebrates - metabolism; Zinc - chemistry
• ISSN: 0961-8368; 1469-896X
• DOI: 10.1002/pro.5560051020

The Cu, Zn superoxide dismutase (Cu, Zn SOD) originally isolated from the periplasmic space of Escherichia coli has been cloned and overexpressed in the E. coli strain BMH 71/18. The protein has been purified as a single component of 17,000 Da, corresponding to one subunit of the common dimeric eukaryotic Cu, Zn SODs. Large crystals of the purified protein have been grown in the presence of polyethylene glycol 4,000 at pH 8.5; the crystals belong to the monoclinic space group P21, with unit cell constants α = 33.1 Å, b = 52.6 Å, c = 43.3 Å, β = 111.4 degrees. One SOD subunit is contained in the asymmetric unit, yielding a Vm value of 2.1 Å3/Da; the crystals diffract X‐rays beyond 2.0 Å resolution.