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Literature summary extracted from

  • Arvizu-Flores, A.A.; Sugich-Miranda, R.; Arreola, R.; Garcia-Orozco, K.D.; Velazquez-Contreras, E.F.; Montfort, W.R.; Maley, F.; Sotelo-Mundo, R.R.
    Role of an invariant lysine residue in folate binding on Escherichia coli thymidylate synthase: calorimetric and crystallographic analysis of the K48Q mutant (2008), Int. J. Biochem. Cell Biol., 40, 2206-2217.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.45 expression in Escherichia coli Escherichia coli

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.1.45 hanging drop method at room temperature, the complexes of mutant TS with dUMP crystallize in the cubic form, binary and ternary complexes with 5NO2dUMP crystallize in the hexagonal form Escherichia coli
2.1.1.45 mutant K48Q, in complex with dUMP or 5-nitro-dUMP, to 2.2 A and 2.7 A resolution, respectively. Binding of dUMP is not impaired in the mutant Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.1.1.45 K48Q 430fold decrease in kcat value, about 30fold increase in Km value for (R)-5,10-methylene-5,6,7,8-tetrahydrofolate. Decrease in binding affinity for folate-like inhibitors Escherichia coli
2.1.1.45 K48Q mutant of Escherichia coli thymidylate synthase Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.1.45 (S)-2-(5(((1,2-dihydro-3-methyl-1-oxobenzo(f)quinazolin-9-yl)methyl)amino)1-oxo-2-isoindolinyl)glutaric acid BW1843U89, U89, distorts the thymidylate synthase active site Escherichia coli
2.1.1.45 10-propargyl-5,8-dideazafolate PDDF, folate-like TS inhibitor Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.1.45 0.05
-
dUMP 10fold higher than wild-type thymidylate synthase Escherichia coli
2.1.1.45 0.05
-
dUMP mutant K48Q Escherichia coli
2.1.1.45 0.3
-
(R)-5,10-methylene-5,6,7,8-tetrahydrofolate 30fold higher than wild-type thymidylate synthase Escherichia coli
2.1.1.45 0.3
-
(R)-5,10-methylenetetrahydrofolate mutant K48Q Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.1.45 5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP Escherichia coli thymidylate synthase is the third enzyme in the methylene tetrahydrofolate cycle besides dihydrofolate reductase and serine hydroxymethyltransferase, in many organisms, thymidylate synthase is the only de novo source for dTMP required for DNA synthesis 7,8-dihydrofolate + dTMP
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.45 Escherichia coli P0A884
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.1.1.45 after purification and precipitation with ammonium sulfate, wild-type and K48Q TS are frozen at -80°C Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.45 (R)-5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
-
Escherichia coli 7,8-dihydrofolate + dTMP
-
?
2.1.1.45 (R)-5,10-methylenetetrahydrofolate + dUMP
-
Escherichia coli dihydrofolate + dTMP
-
?
2.1.1.45 5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP
-
Escherichia coli 7,8-dihydrofolate + dTMP
-
?
2.1.1.45 5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP thymidylate synthase is the third enzyme in the methylene tetrahydrofolate cycle besides dihydrofolate reductase and serine hydroxymethyltransferase, in many organisms, thymidylate synthase is the only de novo source for dTMP required for DNA synthesis Escherichia coli 7,8-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 0.016
-
dUMP mutant K48Q Escherichia coli