Literature summary for 1.3.1.91 extracted from

Chen, M.; Yu, J.; Tanaka, Y.; Tanaka, M.; Tanaka, I.; Yao, M.
Structure of dihydrouridine synthase C (DusC) from Escherichia coli (2013), Acta Crystallogr. Sect. F, 69, 834-838 .

Cloned(Commentary)

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 (Commentary)

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

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

Organism	UniProt	Comment	Textmining
Escherichia coli	P33371	-	-

Purification (Commentary)

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

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	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	tRNA recognition mechanism, overview	Escherichia coli	?	-	?

Subunits

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

Synonyms	Comment	Organism
dihydrouridine synthase C	-	Escherichia coli
DusC	-	Escherichia coli
EcoDusC	-	Escherichia coli

General Information

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