Literature summary extracted from
Moon, J.H.; Park, A.K.; Jang, E.H.; Kim, H.S.; Chi, Y.M.
Crystal structure of a PduO-type ATP: cobalamin adenosyltransferase from Burkholderia thailandensis (2008), Proteins, 72, 1066-1070.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.5.1.17 |
from genomic DNA in pET28a for expression with a C-terminal hexa-His tag in Escherichia coli BL21 (DE3) |
Burkholderia thailandensis |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
2.5.1.17 |
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
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.5.1.17 |
Burkholderia thailandensis |
Q2SZ09 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.5.1.17 |
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
EC Number |
Subunits |
Comment |
Organism |
---|
2.5.1.17 |
trimer |
three molecules in asymmteric unit, oligomerisation mediated by ionic interactions and hydrogen bondings |
Burkholderia thailandensis |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
2.5.1.17 |
ATP: cobalamin adenosyltransferase |
- |
Burkholderia thailandensis |
2.5.1.17 |
PduO |
- |
Burkholderia thailandensis |
2.5.1.17 |
PduO-type ATP: cobalamin adenosyltransferase |
- |
Burkholderia thailandensis |