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Literature summary extracted from

  • Tomoike, F.; Nakagawa, N.; Fukui, K.; Yano, T.; Kuramitsu, S.; Masui, R.
    Indispensable residue for uridine binding in the uridine-cytidine kinase family (2017), Biochem. Biophys. Rep., 11, 93-98 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.48 gene UCK2, recombinant expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Homo sapiens
2.7.1.48 recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta(DE3) Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.48 analysis of cytidine-complex structure of hsUCK2 (PDB ID 1UEJ) Homo sapiens
2.7.1.48 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

EC Number Protein Variants Comment Organism
2.7.1.48 H117Y site-directed mutagenesis, the mutation results in a loss of uridine phosphorylation activity of the enzyme Homo sapiens
2.7.1.48 Y93A site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93C site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93D site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93E site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93F site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93G site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93H site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93I site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93K site-directed mutagenesis, inactive mutant Thermus thermophilus
2.7.1.48 Y93L site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93M site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93N site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93P site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93Q site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93R site-directed mutagenesis, inactive mutant Thermus thermophilus
2.7.1.48 Y93S site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93T site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 Y93V site-directed mutagenesis, the mutation restores the uridine kinase activity that has been lost in the wild-type enzyme Thermus thermophilus
2.7.1.48 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.48 additional information
-
 additional information Michaelis-Menten kinetics in a coupled assay with lactate dehydrogenase and pyruvate kinase Homo sapiens
2.7.1.48 additional information
-
 additional information Michaelis-Menten kinetics in a coupled assay with lactate dehydrogenase and pyruvate kinase Thermus thermophilus
2.7.1.48 0.042
-
 cytidine pH 7.2, 25°C, recombinant wild-type enzyme UCK2 Homo sapiens
2.7.1.48 0.058
-
 uridine pH 7.2, 25°C, recombinant wild-type enzyme UCK2 Homo sapiens
2.7.1.48 0.071
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93W Thermus thermophilus
2.7.1.48 0.072
-
 cytidine pH 7.2, 25°C, recombinant wild-type enzyme ttCK Thermus thermophilus
2.7.1.48 0.1
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93F Thermus thermophilus
2.7.1.48 0.138
-
 cytidine pH 7.2, 25°C, recombinant UCK2 mutant Homo sapiens
2.7.1.48 0.186
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93E Thermus thermophilus
2.7.1.48 0.215
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93C Thermus thermophilus
2.7.1.48 0.22
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93H Thermus thermophilus
2.7.1.48 0.225
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93S Thermus thermophilus
2.7.1.48 0.23
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93Q Thermus thermophilus
2.7.1.48 0.236
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93D Thermus thermophilus
2.7.1.48 0.36
-
 uridine pH 7.2, 25°C, recombinant ttCK mutant Y93H Thermus thermophilus
2.7.1.48 0.383
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93A Thermus thermophilus
2.7.1.48 0.42
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93L Thermus thermophilus
2.7.1.48 0.47
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93N Thermus thermophilus
2.7.1.48 0.555
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93P Thermus thermophilus
2.7.1.48 0.574
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93V Thermus thermophilus
2.7.1.48 0.587
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93T Thermus thermophilus
2.7.1.48 0.62
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93G Thermus thermophilus
2.7.1.48 0.95
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93M Thermus thermophilus
2.7.1.48 0.965
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93I Thermus thermophilus
2.7.1.48 1.6
-
 uridine pH 7.2, 25°C, recombinant ttCK mutant Y93Q Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.48 Mg2+ required Homo sapiens
2.7.1.48 Mg2+ required Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.48 ATP + cytidine Homo sapiens
-
 ADP + CMP
-
 ?
2.7.1.48 ATP + cytidine Thermus thermophilus
-
 ADP + CMP
-
 ?
2.7.1.48 ATP + cytidine Thermus thermophilus DSM 579
-
 ADP + CMP
-
 ?
2.7.1.48 ATP + cytidine Thermus thermophilus ATCC 27634
-
 ADP + CMP
-
 ?
2.7.1.48 ATP + uridine Homo sapiens
-
 ADP + UMP
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.48 Homo sapiens Q9BZX2
-
-
2.7.1.48 Thermus thermophilus Q5SKR5
-
-
2.7.1.48 Thermus thermophilus ATCC 27634 Q5SKR5
-
-
2.7.1.48 Thermus thermophilus DSM 579 Q5SKR5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.48 recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography Homo sapiens
2.7.1.48 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.48 ATP + cytidine
-
 Homo sapiens ADP + CMP
-
 ?
2.7.1.48 ATP + cytidine
-
 Thermus thermophilus ADP + CMP
-
 ?
2.7.1.48 ATP + cytidine
-
 Thermus thermophilus DSM 579 ADP + CMP
-
 ?
2.7.1.48 ATP + cytidine
-
 Thermus thermophilus ATCC 27634 ADP + CMP
-
 ?
2.7.1.48 ATP + uridine
-
 Homo sapiens ADP + UMP
-
 ?
2.7.1.48 ATP + uridine reaction of ttCK enzyme mutants Y93H and Y93Q only, no activity with the wild-type enzyme Thermus thermophilus ADP + UMP
-
 ?
2.7.1.48 ATP + uridine reaction of ttCK enzyme mutants Y93H and Y93Q only, no activity with the wild-type enzyme Thermus thermophilus DSM 579 ADP + UMP
-
 ?
2.7.1.48 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

EC Number Subunits Comment Organism
2.7.1.48 More secondary structure analysis of enzyme ttCK, overview Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
2.7.1.48 hsUCK2
-
 Homo sapiens
2.7.1.48 ttCK
-
 Thermus thermophilus
2.7.1.48 UCK2
-
 Homo sapiens
2.7.1.48 uridine-cytidine kinase 2
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.48 25
-
 assay at Homo sapiens
2.7.1.48 25
-
 assay at Thermus thermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.48 0.4
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93W Thermus thermophilus
2.7.1.48 1.1
-
 uridine pH 7.2, 25°C, recombinant ttCK mutant Y93Q Thermus thermophilus
2.7.1.48 1.4
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93D Thermus thermophilus
2.7.1.48 1.5
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93E Thermus thermophilus
2.7.1.48 1.9
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93C Thermus thermophilus
2.7.1.48 2.8
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93Q Thermus thermophilus
2.7.1.48 2.9
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93G Thermus thermophilus
2.7.1.48 2.9
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93I Thermus thermophilus
2.7.1.48 3.1
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93T Thermus thermophilus
2.7.1.48 3.9
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93V Thermus thermophilus
2.7.1.48 4.1
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93N Thermus thermophilus
2.7.1.48 4.1
-
 cytidine pH 7.2, 25°C, recombinant wild-type enzyme ttCK Thermus thermophilus
2.7.1.48 4.3
-
 cytidine pH 7.2, 25°C, recombinant wild-type enzyme UCK2 Homo sapiens
2.7.1.48 4.5
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93A Thermus thermophilus
2.7.1.48 4.6
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93P Thermus thermophilus
2.7.1.48 5.2
-
 uridine pH 7.2, 25°C, recombinant wild-type enzyme UCK2 Homo sapiens
2.7.1.48 5.9
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93F Thermus thermophilus
2.7.1.48 6.4
-
 cytidine pH 7.2, 25°C, recombinant UCK2 mutant Homo sapiens
2.7.1.48 7.1
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93S Thermus thermophilus
2.7.1.48 7.2
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93M Thermus thermophilus
2.7.1.48 8.4
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93H Thermus thermophilus
2.7.1.48 9.2
-
 uridine pH 7.2, 25°C, recombinant ttCK mutant Y93H Thermus thermophilus
2.7.1.48 10
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93L Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.48 7.2
-
 assay at Homo sapiens
2.7.1.48 7.2
-
 assay at Thermus thermophilus

General Information

EC Number General Information Comment Organism
2.7.1.48 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
2.7.1.48 malfunction mutation H117Y of UCK2 results in a loss of uridine phosphorylation activity of the enzyme Homo sapiens
2.7.1.48 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
2.7.1.48 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
2.7.1.48 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
2.7.1.48 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 [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.1.48 0.689
-
 uridine pH 7.2, 25°C, recombinant ttCK mutant Y93Q Thermus thermophilus
2.7.1.48 4.04
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93I Thermus thermophilus
2.7.1.48 4.68
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93G Thermus thermophilus
2.7.1.48 5.05
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93V Thermus thermophilus
2.7.1.48 5.28
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93T Thermus thermophilus
2.7.1.48 5.63
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93W Thermus thermophilus
2.7.1.48 5.93
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93D Thermus thermophilus
2.7.1.48 7.58
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93M Thermus thermophilus
2.7.1.48 8.07
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93E Thermus thermophilus
2.7.1.48 8.29
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93P Thermus thermophilus
2.7.1.48 8.72
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93N Thermus thermophilus
2.7.1.48 8.84
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93C Thermus thermophilus
2.7.1.48 11.75
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93A Thermus thermophilus
2.7.1.48 12.17
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93Q Thermus thermophilus
2.7.1.48 23.81
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93L Thermus thermophilus
2.7.1.48 25.56
-
 uridine pH 7.2, 25°C, recombinant ttCK mutant Y93H Thermus thermophilus
2.7.1.48 31.56
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93S Thermus thermophilus
2.7.1.48 38.18
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93H Thermus thermophilus
2.7.1.48 46.38
-
 cytidine pH 7.2, 25°C, recombinant UCK2 mutant Homo sapiens
2.7.1.48 56.94
-
 cytidine pH 7.2, 25°C, recombinant wild-type enzyme ttCK Thermus thermophilus
2.7.1.48 59
-
 cytidine pH 7.2, 25°C, recombinant ttCK mutant Y93F Thermus thermophilus
2.7.1.48 89.66
-
 uridine pH 7.2, 25°C, recombinant wild-type enzyme UCK2 Homo sapiens
2.7.1.48 102.38
-
 cytidine pH 7.2, 25°C, recombinant wild-type enzyme UCK2 Homo sapiens