Cloned (Comment) | Organism |
---|---|
from genomic DNA in pET28a for expression with a C-terminal hexa-His tag in Escherichia coli BL21 (DE3) | Burkholderia thailandensis |
Crystallization (Comment) | Organism |
---|---|
native (PDB: 2ZHY, with ordered N-terminal loop and formed active site) and in complex with ATP (PDB: 2ZHZ, substrate-binding cleft is widened and the N-terminal loop swung out and conformational shift of Arg129 side chain upon ATP-binding, similar structure to human and Lactobacillus reuteri PduO except for interaction of NH2 nitrogen atom of Arg11 with gamma-phosphate of ATP), five alpha helix bundle, monomeric subunits almost identical conformations in both structures, crystals: orthothrombic space group C222(1), three monomers in the asymmetric unit, unit cell parameters: a: 52.55/53.91, b: 148.98/148.17, c: 157.35/158.10, microcrystals (from sitting-drop vapour-diffusion at 22°C) scaled up by hanging-drop vapour diffusion, reservoir solution (pH5.7, 22% (w/v) isopropanol, 12% (v/v) PEG4000), PduO-MgATP complex: ATP soaked into native PduO crystals, molecular replacement using PDB: 2G2D as model | Burkholderia thailandensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Burkholderia thailandensis | Q2SZ09 | - |
- |
Purification (Comment) | Organism |
---|---|
from crude lysate by HiTrap Ni2+-chelating affinity chromatography (gradient elution), gel filtration on HiLoad 16/60 Superdex 200 (pH7.9, 4 mM dihiothreitol), concentration to 15 mg/ml by ultrafiltration, yield: 45 mg pure protein per litre culture | Burkholderia thailandensis |
Subunits | Comment | Organism |
---|---|---|
trimer | three molecules in asymmteric unit, oligomerisation mediated by ionic interactions and hydrogen bondings | Burkholderia thailandensis |
Synonyms | Comment | Organism |
---|---|---|
ATP: cobalamin adenosyltransferase | - |
Burkholderia thailandensis |
PduO | - |
Burkholderia thailandensis |
PduO-type ATP: cobalamin adenosyltransferase | - |
Burkholderia thailandensis |