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Literature summary extracted from

  • Lossani, A.; Torti, A.; Gatti, S.; Bruno, A.; Maserati, R.; Wright, G.E.; Focher, F.
    Thymidine kinase and uridine-cytidine kinase from Entamoeba histolytica: cloning, characterization, and search for specific inhibitors (2009), Parasitology, 136, 595-602.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.21 expressed in Escherichia coli Entamoeba histolytica
2.7.1.48 His-tag, expressed in Escherichia coli Entamoeba histolytica

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.21 3'-ethyl-5-methyl-2'-deoxyuridine
-
 Entamoeba histolytica
2.7.1.21 5-iodo-2'-deoxyuridine potent inhibitor Entamoeba histolytica
2.7.1.21 5-propyl-2'-deoxyuridine
-
 Entamoeba histolytica
2.7.1.21 5-trifluoromethyl-2-deoxyuridine potent inhibitor Entamoeba histolytica
2.7.1.21 6-methyluridine
-
 Entamoeba histolytica
2.7.1.21 alpha-D-5-ethyl-2'-deoxyuridine
-
 Entamoeba histolytica
2.7.1.21 alpha-L-thymidine
-
 Entamoeba histolytica
2.7.1.21 beta-D-5-ethyl-2'-deoxyuridine potent inhibitor Entamoeba histolytica
2.7.1.21 beta-L-thymidine potent inhibitor Entamoeba histolytica
2.7.1.21 additional information not inhibited by D-5-(bromovinyl)-2'-deoxyuridine, L-5-(bromovinyl)-2'-deoxyuridine, and 2-phenylamino-9-(4-hydroxybutyl)-6-oxopurine Entamoeba histolytica
2.7.1.21 penciclovir
-
 Entamoeba histolytica
2.7.1.48 5-propyl-2'-deoxyuridine
-
 Entamoeba histolytica
2.7.1.48 6-(4-hexyloxyanilino)uracil
-
 Entamoeba histolytica
2.7.1.48 cytidine competitive inhibitor of phosphorylation of uridine Entamoeba histolytica
2.7.1.48 KCl
-
 Entamoeba histolytica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.21 0.00027
-
 thymidine at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica
2.7.1.48 0.22
-
 cytidine
-
 Entamoeba histolytica
2.7.1.48 0.74
-
 uridine
-
 Entamoeba histolytica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.21 Mg2+ MgCl2 is required for activity, reaching its optimal effect at 1.2 mM when ATP is present at 1 mM Entamoeba histolytica
2.7.1.21 Mn2+ MnCl2 can replace MgCl2 in the assay, with optimal activity at 200 mM when ATP is present at 1 mM Entamoeba histolytica
2.7.1.48 Mg2+
-
 Entamoeba histolytica

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.48 40000
-
 x * 40000 Da, SDS-PAGE Entamoeba histolytica

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.21 Entamoeba histolytica Q6S4W2
-
-
2.7.1.48 Entamoeba histolytica C4M4T4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.21 Ni-NTA column chromatography Entamoeba histolytica
2.7.1.48 immobilized metal ion adsorption chromatography (Ni2+) Entamoeba histolytica

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.48 0.17
-
-
 Entamoeba histolytica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.21 ATP + 5-iodo-2'-deoxyuridine 5-iodo-2'-deoxyuridine is efficiently phosphorylated by thymidine kinase confirming that these compound enters the active site of the enzyme and acts as alternate substrate Entamoeba histolytica ?
-
 ?
2.7.1.21 ATP + 5-trifluoromethyl-2-deoxyuridine 5-trifluoromethyl-2-deoxyuridine is efficiently phosphorylated by thymidine kinase confirming that these compound enters the active site of the enzyme and acts as alternate substrate Entamoeba histolytica ?
-
 ?
2.7.1.21 ATP + beta-5-ethyl-2'-deoxyuridine beta-5-ethyl-2'-deoxyuridine is efficiently phosphorylated by thymidine kinase confirming that these compound enters the active site of the enzyme and acts as alternate substrate Entamoeba histolytica ?
-
 ?
2.7.1.21 ATP + thymidine ATP acts as specific phosphate donor Entamoeba histolytica ADP + thymidine 5'-phosphate
-
 ?
2.7.1.21 additional information beta-L-thymidine is poorly phosphorylated by the enzyme Entamoeba histolytica ?
-
 ?
2.7.1.21 additional information does not use UTP, GTP, and TTP as cosubstrates Entamoeba histolytica ?
-
 ?
2.7.1.48 cytidine + ATP
-
 Entamoeba histolytica CMP + ADP
-
 ?
2.7.1.48 uridine + ATP with GTP: 25% activity, with UTP or CTP: 0% activity Entamoeba histolytica UMP + ADP
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.48 ? x * 40000 Da, SDS-PAGE Entamoeba histolytica

Synonyms

EC Number Synonyms Comment Organism
2.7.1.48 uridine-cytidine kinase
-
 Entamoeba histolytica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.21 8
-
 exhibits best activity at pH 8.0 at an ATP concentration range between 0.2 and 4.0 mM Entamoeba histolytica
2.7.1.48 7.6
-
-
 Entamoeba histolytica

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.21 ATP
-
 Entamoeba histolytica

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.7.1.21 0.0006
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica 5-trifluoromethyl-2-deoxyuridine
2.7.1.21 0.001
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica 5-iodo-2'-deoxyuridine
2.7.1.21 0.0025
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica beta-D-5-ethyl-2'-deoxyuridine
2.7.1.21 0.005
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica beta-L-thymidine
2.7.1.21 0.01
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica penciclovir
2.7.1.21 0.015
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica 6-methyluridine
2.7.1.21 0.017
-
-
 Entamoeba histolytica alpha-D-5-ethyl-2'-deoxyuridine
2.7.1.21 0.018
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica 3'-ethyl-5-methyl-2'-deoxyuridine
2.7.1.21 0.035
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica alpha-L-thymidine
2.7.1.21 0.0444
-
 at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica 5-propyl-2'-deoxyuridine
2.7.1.48 0.054
-
-
 Entamoeba histolytica 6-(4-hexyloxyanilino)uracil
2.7.1.48 0.09
-
-
 Entamoeba histolytica 5-propyl-2'-deoxyuridine
2.7.1.48 250
-
-
 Entamoeba histolytica KCl