Literature summary for 3.5.4.13 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.

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
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
R115Q	site-directed mutagenesis, inactive mutant	Escherichia coli
S111C	site-directed mutagenesis, inactive mutant	Escherichia coli
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

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

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Escherichia coli

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P28248	-	-

Substrates and Products (Substrate)

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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