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Literature summary extracted from

  • Cardinale, D.; Salo-Ahen, O.; Guaitoli, G.; Ferrari, S.; Venturelli, A.; Franchini, S.; Battini, R.; Ponterini, G.; Wade, R.; Costi, M.
    Design and characterization of a mutation outside the active site of human thymidylate synthase that affects ligand binding (2010), Protein Eng., 23, 81-89.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.45 expressed in Escherichia coli BL21 strain DE3pLysS Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.45 E30W the mutant shows 100times lower specific activity with respect to the wild type enzyme and is resistant to 5-fluoro-2'-deoxyuridine-5'-monophosphate (6fold higher inhibition constant than the wild type enzyme) Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.45 5-fluoro-2'-deoxyuridine-5'-monophosphate
-
Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.45 0.0015
-
5,10-methylenetetrahydrofolate wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 0.0054
-
dUMP mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 0.0078
-
dUMP wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 0.0524
-
5,10-methylenetetrahydrofolate mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.45 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.45
-
Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.45 5,10-methylenetetrahydrofolate + dUMP
-
Homo sapiens dihydrofolate + dTMP
-
?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.45 3.9
-
5,10-methylenetetrahydrofolate mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 3.9
-
dUMP mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 48
-
5,10-methylenetetrahydrofolate wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 48
-
dUMP wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.1.1.45 0.0011
-
5-fluoro-2'-deoxyuridine-5'-monophosphate wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 0.0054
-
5-fluoro-2'-deoxyuridine-5'-monophosphate mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.45 70
-
5,10-methylenetetrahydrofolate mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 720
-
dUMP mutant enzyme E30W, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 3200
-
5,10-methylenetetrahydrofolate wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens
2.1.1.45 6150
-
dUMP wild type enzyme, in 50 mM TES buffer, 50 mM, MgCl2, 13 mM formalin, 2 mM EDTA, 0.15 mM 2-mercaptoethanol, pH 7.4, at 25°C Homo sapiens