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Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of GK0767, the copper-containing nitrite reductase from Geobacillus kaustophilus.

ABSTRACT: The soluble region (residues 32-354) of GK0767, a copper-containing nitrite reductase from the thermophilic Gram-positive bacterium Geobacillus kaustophilus HTA426, has been cloned and overexpressed in Escherichia coli. The purified recombinant protein was crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected and processed to a maximum resolution of 1.3?Å. The crystals belonged to space group R3, with unit-cell parameters a = b = 115.1, c = 87.5?Å. Preliminary studies and molecular-replacement calculations reveal the presence of one subunit of the homotrimeric structure in the asymmetric unit; this corresponds to a V(M) value of 3.14?Å(3)?Da(-1).

SUBMITTER: Fukuda Y

PROVIDER: S-EPMC3107145 | biostudies-literature | 2011 Jun

REPOSITORIES: biostudies-literature

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Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of GK0767, the copper-containing nitrite reductase from Geobacillus kaustophilus.

Fukuda Yohta Y Tamada Taro T Takami Hideto H Suzuki Shinnichiro S Inoue Tsuyoshi T Nojiri Masaki M

Acta crystallographica. Section F, Structural biology and crystallization communications 20110526 Pt 6

The soluble region (residues 32-354) of GK0767, a copper-containing nitrite reductase from the thermophilic Gram-positive bacterium Geobacillus kaustophilus HTA426, has been cloned and overexpressed in Escherichia coli. The purified recombinant protein was crystallized using the hanging-drop vapour-diffusion method. X-ray diffraction data were collected and processed to a maximum resolution of 1.3 Å. The crystals belonged to space group R3, with unit-cell parameters a = b = 115.1, c = 87.5 Å. Pr ...[more]

PMID: 21636914

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