Open AccessCloning, expression, purification, crystallization and preliminary X‐ray crystallographic study of GK0767, the copper‐containing nitrite reductase from Geobacillus kaustophilus
Author(s) -
Fukuda Yohta,
Tamada Taro,
Takami Hideto,
Suzuki Shinnichiro,
Inoue Tsuyoshi,
Nojiri Masaki
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111013297
Subject(s) - thermophile , escherichia coli , nitrite reductase , crystallization , protein subunit , chemistry , crystallography , molecular replacement , recombinant dna , biochemistry , reductase , stereochemistry , enzyme , nitrate reductase , crystal structure , organic chemistry , gene
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 R 3, 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 .