Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli strain BL21-CodonPlus(DE3)-RIL | Pyrococcus horikoshii |
Crystallization (Comment) | Organism |
---|---|
enzyme with a phosphate ion occupying the position of the beta-phosphate of the nucleotide, mixing of 0.001 ml of protein solution containing 1.95 mg/ml protein in 20 mM Tris-HCl containing 200 mM NaCl and 1 mM DTT, pH 8.0, with 0.001 ml of reservoir solution containing 0.1 M CHESS buffer, pH 9.3, and 0.88 M sodium citrate as the precipitant, 20°C, 1 week, 25% v/v glycerol for cryoprotection, X-ray diffraction structure determination and analysis at 1.85 A resolution | Pyrococcus horikoshii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | - |
- |
- |
Pyrococcus horikoshii OT-3 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIL by gel filtration, anion exchange chromatography to over 95% purity | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the active site is located at the interface between adjacent subunits. Each active site contains a thiazole-binding site and a nucleotide-binding site, structure, overview. Asp residue as a catalytic base | Pyrococcus horikoshii | ? | - |
? | |
additional information | the active site is located at the interface between adjacent subunits. Each active site contains a thiazole-binding site and a nucleotide-binding site, structure, overview. Asp residue as a catalytic base | Pyrococcus horikoshii OT-3 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | monomer structure, docking of ternary complexes, tertiary structure, molecular modelling, overview | Pyrococcus horikoshii |
trimer | residues involved in hydrogen bonds between subunits in the enzyme trimer, overview | Pyrococcus horikoshii |
Synonyms | Comment | Organism |
---|---|---|
4-methyl-5-beta-hydroxyethylthiazole kinase | - |
Pyrococcus horikoshii |
ThiK | - |
Pyrococcus horikoshii |
General Information | Comment | Organism |
---|---|---|
evolution | three-quarters of the residues involved in interfacial regions are conserved, also the amino-acid residues in the nucleotide-binding, magnesium ion-binding and substrate-binding sites are conserved. The overall structure of PhThiK is similar to those of Bacillus subtilis ThiK (BsThiK) and Enterococcus faecalis V583 ThiK (EfThiK) | Pyrococcus horikoshii |
additional information | the enzyme topology shows the typical ribokinase fold of an alpha/beta protein. Binding of the nucleotide and substrate to the ThiK enzyme do not influence the trimeric quaternary association | Pyrococcus horikoshii |