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Literature summary for 2.5.1.17 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.
    View publication on PubMed

Cloned(Commentary)

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 (Commentary)

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

Organism UniProt Comment Textmining
Burkholderia thailandensis Q2SZ09
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Purification (Commentary)

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

Subunits Comment Organism
trimer three molecules in asymmteric unit, oligomerisation mediated by ionic interactions and hydrogen bondings Burkholderia thailandensis

Synonyms

Synonyms Comment Organism
ATP: cobalamin adenosyltransferase
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Burkholderia thailandensis
PduO
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Burkholderia thailandensis
PduO-type ATP: cobalamin adenosyltransferase
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Burkholderia thailandensis