Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme as wild-type and selenomethionine-labeled proteins in Escherichia coli strains BL21(DE3) and B834 (DE3), respectively | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged and SeMet-labeled DusC, sitting drop vapour diffusion method, mixing of 200 nl of about 10 mg/ml of protein in 20 mM HEPES, pH 7.6, 1 mM MgCl2, 200 mM KCl, 7 mM 2-mercaptoethanol, and 10% glycerol, with 200 nl of reservoir solution containing 0.1 M Tris, pH 7.9, 0.2 M sodium acetate, and 12% PEG 4000 for the wild-type enzyme and 0.1 M imidazole, pH 8.0, 15% v/v 2-propanol, and 20% v/v glycerol for the SeMet-labeled enzyme, X-ray diffraction structure determination and analysis at 2.1 A resolution | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P33371 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and selenomethionine-labeled enzymes from Escherichia coli cell-free extract (centrifugation at 40000 x g) by nickel affinity chromatography, dialysis, heparin affinity chromatography, and gel filtration | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
5,6-dihydrouracil20 in tRNA + NAD(P)+ = uracil20 in tRNA + NAD(P)H + H+ | Dus proteins adopt a common substrate recognition mechanism using an adapter molecule, whereas the manner of tRNA binding is diverse | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | tRNA recognition mechanism, overview | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme is composed of two domains: an N-terminal catalytic domain and a C-terminal tRNA-binding domain | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
dihydrouridine synthase C | - |
Escherichia coli |
DusC | - |
Escherichia coli |
EcoDusC | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | comparison of structure and substrate recognition mechanisms of Thermus thermophilus Dus and Escherichia coli Dus enzymes, overview | Escherichia coli |
additional information | residues that participate in binding to the adapter molecule in EcoDusC are Asn95, Lys139, Arg141, His168 and Arg170. The catalytic cysteine residue is Cys98 | Escherichia coli |