Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme, functional complementation of an enzyme-deficient Saccharomyces cerevisiae mutant by expression of enzyme domains HsDus2dusD and HsDus2dsRBD, both domains are required for activity | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant catalytic HsDus2dusD domain and tRNA binding domain HsDus2dsRBD (Glu347-Lys451), by hanging drop vapor diffusion method, at 19°C, X-ray diffraction structure determination and crystal structure analysis, PDB IDs 4WFS and 4WFT | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
uracil20 in tRNA + NAD(P)H + H+ | Homo sapiens | - |
5,6-dihydrouracil20 in tRNA + NAD(P)+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9NX74 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme by nickel affinity chromatography and gel filtration | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
5,6-dihydrouracil20 in tRNA + NAD(P)+ = uracil20 in tRNA + NAD(P)H + H+ | dsRNA binding mode, modeling | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
lung cancer cell | isozyme Dus2 is upregulated | Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | no enzymatic activity ith tRNAAsp substrate | Homo sapiens | ? | - |
? | |
uracil20 in tRNA + NAD(P)H + H+ | - |
Homo sapiens | 5,6-dihydrouracil20 in tRNA + NAD(P)+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 55000, about, recombinant enzyme, SDS-PAGE | Homo sapiens |
More | the N-terminal catalytic domain contains the flavin cofactor involved in the reduction of uridine. The second module is the conserved alpha-helical domain known as the tRNA binding domain in HsDus2 homologues. It is connected via a flexible linker to an unusual extended version of a dsRNA binding domain (dsRBD). The catalytic domain binds selectively NADPH but cannot reduce uridine in the absence of the dsRBD. Domain architecture of HsDus2, enzyme MALDI peptide mass finger printing analysis, overview | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
dihydrouridine synthase 2 | - |
Homo sapiens |
DUS2 | - |
Homo sapiens |
HsDus2 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FMN | the flavin cofactor is involved in the reduction of uridine, FMN binding site structure, overview. HsDus2dusD binds the FMN cofactor non-covalently at the C-terminal end of the beta-barrel, above beta-strands 1 and 8 | Homo sapiens | |
NAD(P)H | binds to the catalytic domain | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
evolution | while in Dus enzymes from bacteria, plants and fungi, tRNA binding is essentially achieved by the alpha-helical domain, in HsDus2 this function is carried out by the dsRNA binding domain (dsRBD) | Homo sapiens |
additional information | the catalytic domain binds selectively NADPH but cannot reduce uridine in the absence of the dsRNA binding domain (dsRBD). HsDus2 catalytic domain structure, overview | Homo sapiens |
physiological function | in tRNA, dihydrouridine is a conserved modified base generated by the post-transcriptional reduction of uridine. Formation of dihydrouridine 20, located in the D-loop, is catalyzed by dihydrouridine synthase 2 (Dus2). Full-length HsDus2 but not its Dus domain complements a DELTAdus2 yeast strain by catalyzing formation of dihydrouridine20 | Homo sapiens |