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Literature summary for 2.1.1.45 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)

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

Protein Variants

Protein Variants Comment Organism
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

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Homo sapiens

Substrates and Products (Substrate)

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
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
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]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
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
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
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