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Literature summary for 2.7.1.21 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)

Cloned (Comment) Organism
expressed in Escherichia coli Entamoeba histolytica

Inhibitors

Inhibitors Comment Organism Structure
3'-ethyl-5-methyl-2'-deoxyuridine
-
Entamoeba histolytica
5-iodo-2'-deoxyuridine potent inhibitor Entamoeba histolytica
5-propyl-2'-deoxyuridine
-
Entamoeba histolytica
5-trifluoromethyl-2-deoxyuridine potent inhibitor Entamoeba histolytica
6-methyluridine
-
Entamoeba histolytica
alpha-D-5-ethyl-2'-deoxyuridine
-
Entamoeba histolytica
alpha-L-thymidine
-
Entamoeba histolytica
beta-D-5-ethyl-2'-deoxyuridine potent inhibitor Entamoeba histolytica
beta-L-thymidine potent inhibitor Entamoeba histolytica
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
penciclovir
-
Entamoeba histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00027
-
thymidine at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
Entamoeba histolytica Q6S4W2
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Entamoeba histolytica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 ?
-
?
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 ?
-
?
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 ?
-
?
ATP + thymidine ATP acts as specific phosphate donor Entamoeba histolytica ADP + thymidine 5'-phosphate
-
?
additional information beta-L-thymidine is poorly phosphorylated by the enzyme Entamoeba histolytica ?
-
?
additional information does not use UTP, GTP, and TTP as cosubstrates Entamoeba histolytica ?
-
?

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
Entamoeba histolytica

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
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
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
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
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
0.01
-
at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica penciclovir
0.015
-
at 37°C for 20 min in 30 mM potassium phosphate, pH 8.0, 1.2 mM MgCl2 Entamoeba histolytica 6-methyluridine
0.017
-
-
Entamoeba histolytica alpha-D-5-ethyl-2'-deoxyuridine
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
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
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