Computational protocol: Crystal Structure of a Nitrate/Nitrite Exchanger

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Protocol publication

[…] NarK from E. coli strain K12 was overexpressed in E. coli BL21 (DE3) C41. Fab antibody fragments were generated as described in Methods. NarK-Fab complex was purified in the presence of 0.2 % (w/v) n-decyl β-D-maltoside and crystallized in the following condition: 0.1M citric acid (pH 3.5), 0.1M NaCl, 0.1M Li2SO4 and 28% PEG400. Nitrite-bound NarK-Fab crystal was obtained by soaking the NarK-Fab crystal in the buffer containing 50mM sodium nitrite. Diffraction data sets of both crystals were collected at the Advanced Light Source (beamline 8.2.2). Data processing and structure determination were performed using the HKL2000, COOT and CCP4 programs. […]

Pipeline specifications

Software tools Coot, CCP4
Application Protein structure analysis
Organisms Escherichia coli
Chemicals Nitrates, Nitric Oxide, Nitrites, Ammonium Compounds