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Information on EC 1.3.1.91 - tRNA-dihydrouridine20 synthase [NAD(P)+] and Organism(s) Escherichia coli and UniProt Accession P33371

for references in articles please use BRENDA:EC1.3.1.91
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IUBMB Comments
A flavoenzyme . The enzyme specifically modifies uracil20 in tRNA.
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This record set is specific for:
Escherichia coli
UNIPROT: P33371
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Word Map
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Reaction Schemes
5,6-dihydrouracil20 in tRNA
+
=
uracil20 in tRNA
+
+
Synonyms
hdus2, dihydrouridine synthase, hsdus2, dihydrouridine synthase 2, dus 2, dus2p, ecodusc, trna-dihydrouridine synthases, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dihydrouridine synthase C
-
tRNA-dihydrouridine synthases
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
5,6-dihydrouracil20 in tRNA + NAD(P)+ = uracil20 in tRNA + NAD(P)H + H+
show the reaction diagram
Dus proteins adopt a common substrate recognition mechanism using an adapter molecule, whereas the manner of tRNA binding is diverse
SYSTEMATIC NAME
IUBMB Comments
tRNA-5,6-dihydrouracil20:NAD(P)+ oxidoreductase
A flavoenzyme [3]. The enzyme specifically modifies uracil20 in tRNA.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
uracil20 in tRNA + NADH + H+
5,6-dihydrouracil20 in tRNA + NAD+
show the reaction diagram
-
-
-
?
uracil20a in tRNA + NADH + H+
5,6-dihydrouracil20a in tRNA + NAD+
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
uracil20 in tRNA + NADH + H+
5,6-dihydrouracil20 in tRNA + NAD+
show the reaction diagram
-
-
-
?
uracil20a in tRNA + NADH + H+
5,6-dihydrouracil20a in tRNA + NAD+
show the reaction diagram
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
2 mM used in assay conditions
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
comparison of structure and substrate recognition mechanisms of Thermus thermophilus Dus and Escherichia coli Dus enzymes, overview
additional information
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
vapor diffusion method, using 1.6 M lithium sulfate and 100 mM HEPES pH 7.5
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
Ni-NTA column chromatography and Superdex 200 gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant expression of His-tagged enzyme as wild-type and selenomethionine-labeled proteins in Escherichia coli strains BL21(DE3) and B834 (DE3), respectively
expressed in Escherichia coli BL21(DE3) cells
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Chen, M.; Yu, J.; Tanaka, Y.; Tanaka, M.; Tanaka, I.; Yao, M.
Structure of dihydrouridine synthase C (DusC) from Escherichia coli
Acta Crystallogr. Sect. F
69
834-838
2013
Escherichia coli (P33371), Escherichia coli
Manually annotated by BRENDA team
Bou-Nader, C.; Montemont, H.; Guerineau, V.; Jean-Jean, O.; Bregeon, D.; Hamdane, D.
Unveiling structural and functional divergences of bacterial tRNA dihydrouridine synthases Perspectives on the evolution scenario
Nucleic Acids Res.
46
1386-1394
2018
Escherichia coli (P32695)
Manually annotated by BRENDA team