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Literature summary extracted from

  • Thymark, M.; Johansson, E.; Larsen, S.; Willemoes, M.
    Mutational analysis of the nucleotide binding site of Escherichia coli dCTP deaminase (2008), Arch. Biochem. Biophys., 470, 20-26.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.4.13 recombinant mutant E138D in complex with dUTP, hanging drop vapour diffusion method, 0.002 ml of 2 mg/ml protein in solution with 5 mM dUTP and 20 mM MgSO4 is mixed with 0.002 ml reservoir solution containing 27.5% PEG 400, 50 mM MgSO4 and 0.1 M HEPES, pH 7.5, equilibration against 0.5 ml of reservoir solution, room temeprature, crystal X-ray diffraction structure determination and analysis at 2.1 A resolution Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
3.5.4.13 E138D site-directed mutagenesis, the mutant enzyme shows a 140fold reduction in kcat and altered dTTP inhibition compared to the wild-type enzyme Escherichia coli
3.5.4.13 R115Q site-directed mutagenesis, inactive mutant Escherichia coli
3.5.4.13 S111C site-directed mutagenesis, inactive mutant Escherichia coli
3.5.4.13 S111T site-directed mutagenesis, the mutant enzyme shows a 30fold reduction in kcat and altered dTTP inhibition compared to the wild-type enzyme, modeling of the active site of the S111T enzyme, overview Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.4.13 dTTP inhibits by increasing the kcat several fold, mutants S111T and E138D are less sensitive to dTTP, overview Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.4.13 Mg2+
-
 Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.4.13 dCTP + H2O Escherichia coli
-
 dUTP + NH3 dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.4.13 Escherichia coli P28248
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.4.13 dCTP + H2O
-
 Escherichia coli dUTP + NH3
-
 ?
3.5.4.13 dCTP + H2O
-
 Escherichia coli dUTP + NH3 dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.4.13 37
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.4.13 additional information
-
 additional information
-
 Escherichia coli
3.5.4.13 1.24
-
 dCTP pH 6.8, 37°C, recombinant wild-type enzyme Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.4.13 6.8
-
 assay at Escherichia coli