Cloned (Comment) | Organism |
---|---|
gene UCK2, recombinant expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Homo sapiens |
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta(DE3) | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
analysis of cytidine-complex structure of hsUCK2 (PDB ID 1UEJ) | Homo sapiens |
purified recombinant Y93H ttCK in complex with cytidine and AMPPCP, X-ray diffraction structure determination and analysis at 2.49 A resolution | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
H117Y | site-directed mutagenesis, the mutation results in a loss of uridine phosphorylation activity of the enzyme | Homo sapiens |
Y93A | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93C | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93D | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93E | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93F | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93G | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93H | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93I | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93K | site-directed mutagenesis, inactive mutant | Thermus thermophilus |
Y93L | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93M | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93N | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93P | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93Q | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93R | site-directed mutagenesis, inactive mutant | Thermus thermophilus |
Y93S | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93T | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93V | site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
Y93W | site-directed mutagenesis, the mutation does not restore the uridine kinase activity that has been lost in the wild-type enzyme | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics in a coupled assay with lactate dehydrogenase and pyruvate kinase | Homo sapiens | |
additional information | - |
additional information | Michaelis-Menten kinetics in a coupled assay with lactate dehydrogenase and pyruvate kinase | Thermus thermophilus | |
0.042 | - |
cytidine | pH 7.2, 25°C, recombinant wild-type enzyme UCK2 | Homo sapiens | |
0.058 | - |
uridine | pH 7.2, 25°C, recombinant wild-type enzyme UCK2 | Homo sapiens | |
0.071 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93W | Thermus thermophilus | |
0.072 | - |
cytidine | pH 7.2, 25°C, recombinant wild-type enzyme ttCK | Thermus thermophilus | |
0.1 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93F | Thermus thermophilus | |
0.138 | - |
cytidine | pH 7.2, 25°C, recombinant UCK2 mutant | Homo sapiens | |
0.186 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93E | Thermus thermophilus | |
0.215 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93C | Thermus thermophilus | |
0.22 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93H | Thermus thermophilus | |
0.225 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93S | Thermus thermophilus | |
0.23 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93Q | Thermus thermophilus | |
0.236 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93D | Thermus thermophilus | |
0.36 | - |
uridine | pH 7.2, 25°C, recombinant ttCK mutant Y93H | Thermus thermophilus | |
0.383 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93A | Thermus thermophilus | |
0.42 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93L | Thermus thermophilus | |
0.47 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93N | Thermus thermophilus | |
0.555 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93P | Thermus thermophilus | |
0.574 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93V | Thermus thermophilus | |
0.587 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93T | Thermus thermophilus | |
0.62 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93G | Thermus thermophilus | |
0.95 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93M | Thermus thermophilus | |
0.965 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93I | Thermus thermophilus | |
1.6 | - |
uridine | pH 7.2, 25°C, recombinant ttCK mutant Y93Q | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Homo sapiens | |
Mg2+ | required | Thermus thermophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + cytidine | Homo sapiens | - |
ADP + CMP | - |
? | |
ATP + cytidine | Thermus thermophilus | - |
ADP + CMP | - |
? | |
ATP + cytidine | Thermus thermophilus DSM 579 | - |
ADP + CMP | - |
? | |
ATP + cytidine | Thermus thermophilus ATCC 27634 | - |
ADP + CMP | - |
? | |
ATP + uridine | Homo sapiens | - |
ADP + UMP | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9BZX2 | - |
- |
Thermus thermophilus | Q5SKR5 | - |
- |
Thermus thermophilus ATCC 27634 | Q5SKR5 | - |
- |
Thermus thermophilus DSM 579 | Q5SKR5 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography | Homo sapiens |
recombinant wild-type and mutant enzymes from Escherichia coli strain Rosetta(DE3) by heat treatment at 70°C for 20 min, followed by hydrophobic interaction chromatography, and two different steps of gel filtration | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + cytidine | - |
Homo sapiens | ADP + CMP | - |
? | |
ATP + cytidine | - |
Thermus thermophilus | ADP + CMP | - |
? | |
ATP + cytidine | - |
Thermus thermophilus DSM 579 | ADP + CMP | - |
? | |
ATP + cytidine | - |
Thermus thermophilus ATCC 27634 | ADP + CMP | - |
? | |
ATP + uridine | - |
Homo sapiens | ADP + UMP | - |
? | |
ATP + uridine | reaction of ttCK enzyme mutants Y93H and Y93Q only, no activity with the wild-type enzyme | Thermus thermophilus | ADP + UMP | - |
? | |
ATP + uridine | reaction of ttCK enzyme mutants Y93H and Y93Q only, no activity with the wild-type enzyme | Thermus thermophilus DSM 579 | ADP + UMP | - |
? | |
ATP + uridine | reaction of ttCK enzyme mutants Y93H and Y93Q only, no activity with the wild-type enzyme | Thermus thermophilus ATCC 27634 | ADP + UMP | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | secondary structure analysis of enzyme ttCK, overview | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
hsUCK2 | - |
Homo sapiens |
ttCK | - |
Thermus thermophilus |
UCK2 | - |
Homo sapiens |
uridine-cytidine kinase 2 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
25 | - |
assay at | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.4 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93W | Thermus thermophilus | |
1.1 | - |
uridine | pH 7.2, 25°C, recombinant ttCK mutant Y93Q | Thermus thermophilus | |
1.4 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93D | Thermus thermophilus | |
1.5 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93E | Thermus thermophilus | |
1.9 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93C | Thermus thermophilus | |
2.8 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93Q | Thermus thermophilus | |
2.9 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93G | Thermus thermophilus | |
2.9 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93I | Thermus thermophilus | |
3.1 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93T | Thermus thermophilus | |
3.9 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93V | Thermus thermophilus | |
4.1 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93N | Thermus thermophilus | |
4.1 | - |
cytidine | pH 7.2, 25°C, recombinant wild-type enzyme ttCK | Thermus thermophilus | |
4.3 | - |
cytidine | pH 7.2, 25°C, recombinant wild-type enzyme UCK2 | Homo sapiens | |
4.5 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93A | Thermus thermophilus | |
4.6 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93P | Thermus thermophilus | |
5.2 | - |
uridine | pH 7.2, 25°C, recombinant wild-type enzyme UCK2 | Homo sapiens | |
5.9 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93F | Thermus thermophilus | |
6.4 | - |
cytidine | pH 7.2, 25°C, recombinant UCK2 mutant | Homo sapiens | |
7.1 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93S | Thermus thermophilus | |
7.2 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93M | Thermus thermophilus | |
8.4 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93H | Thermus thermophilus | |
9.2 | - |
uridine | pH 7.2, 25°C, recombinant ttCK mutant Y93H | Thermus thermophilus | |
10 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93L | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Homo sapiens |
7.2 | - |
assay at | Thermus thermophilus |
General Information | Comment | Organism |
---|---|---|
malfunction | mutagenesis of Tyr93 in ttCK reveals that the uridine phosphorylation activity is restored only by replacement of Tyr93 with His or Gln | Thermus thermophilus |
malfunction | mutation H117Y of UCK2 results in a loss of uridine phosphorylation activity of the enzyme | Homo sapiens |
additional information | uridine phosphorylation activity commonly depends on a single residue in the UCK family. Molecular docking analysis. Structure comparison of Thermus thermophilus enzyme ttCK and human enzyme hsUCK2 | Thermus thermophilus |
additional information | uridine phosphorylation activity commonly depends on a single residue in the UCK family. Structure comparison of Thermus thermophilus enzyme ttCK and human enzyme hsUCK2 | Homo sapiens |
physiological function | uridine-cytidine kinase (UCK) is one of the enzymes in the nucleoside salvage pathway. UCK generally converts both cytidine and uridine to nucleoside monophosphate using ATP as the phosphate donor | Homo sapiens |
physiological function | uridine-cytidine kinase (UCK) is one of the enzymes in the nucleoside salvage pathway. UCK generally converts both cytidine and uridine to nucleoside monophosphate using ATP as the phosphate donor, but the UCK of Thermus thermophilus HB8 (ttCK) phosphorylates only cytidine. This cytidine-restricted activity is thought to depend on residue Tyr93 | Thermus thermophilus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.689 | - |
uridine | pH 7.2, 25°C, recombinant ttCK mutant Y93Q | Thermus thermophilus | |
4.04 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93I | Thermus thermophilus | |
4.68 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93G | Thermus thermophilus | |
5.05 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93V | Thermus thermophilus | |
5.28 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93T | Thermus thermophilus | |
5.63 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93W | Thermus thermophilus | |
5.93 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93D | Thermus thermophilus | |
7.58 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93M | Thermus thermophilus | |
8.07 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93E | Thermus thermophilus | |
8.29 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93P | Thermus thermophilus | |
8.72 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93N | Thermus thermophilus | |
8.84 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93C | Thermus thermophilus | |
11.75 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93A | Thermus thermophilus | |
12.17 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93Q | Thermus thermophilus | |
23.81 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93L | Thermus thermophilus | |
25.56 | - |
uridine | pH 7.2, 25°C, recombinant ttCK mutant Y93H | Thermus thermophilus | |
31.56 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93S | Thermus thermophilus | |
38.18 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93H | Thermus thermophilus | |
46.38 | - |
cytidine | pH 7.2, 25°C, recombinant UCK2 mutant | Homo sapiens | |
56.94 | - |
cytidine | pH 7.2, 25°C, recombinant wild-type enzyme ttCK | Thermus thermophilus | |
59 | - |
cytidine | pH 7.2, 25°C, recombinant ttCK mutant Y93F | Thermus thermophilus | |
89.66 | - |
uridine | pH 7.2, 25°C, recombinant wild-type enzyme UCK2 | Homo sapiens | |
102.38 | - |
cytidine | pH 7.2, 25°C, recombinant wild-type enzyme UCK2 | Homo sapiens |