BRENDA - Enzyme Database show
show all sequences of 3.5.4.13

Mutational analysis of the nucleotide binding site of Escherichia coli dCTP deaminase

Thymark, M.; Johansson, E.; Larsen, S.; Willemoes, M.; Arch. Biochem. Biophys. 470, 20-26 (2008)

Data extracted from this reference:

Crystallization (Commentary)
Crystallization
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
Engineering
Amino acid exchange
Commentary
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
Commentary
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
Commentary
Organism
Structure
Mg2+
-
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
dCTP + H2O
Escherichia coli
-
dUTP + NH3
dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P28248
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dCTP + H2O
-
684700
Escherichia coli
dUTP + NH3
-
-
-
?
dCTP + H2O
-
684700
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]
Commentary
Organism
37
-
assay at
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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
Commentary
Organism
6.8
-
assay at
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
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
Engineering (protein specific)
Amino acid exchange
Commentary
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 (protein specific)
Inhibitors
Commentary
Organism
Structure
dTTP
inhibits by increasing the kcat several fold, mutants S111T and E138D are less sensitive to dTTP, overview
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
-
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
dCTP + H2O
Escherichia coli
-
dUTP + NH3
dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
dCTP + H2O
-
684700
Escherichia coli
dUTP + NH3
-
-
-
?
dCTP + H2O
-
684700
Escherichia coli
dUTP + NH3
dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
-
Escherichia coli
1.24
-
dCTP
pH 6.8, 37°C, recombinant wild-type enzyme
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.8
-
assay at
Escherichia coli
Other publictions for EC 3.5.4.13
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733177
Oehlenschlaeger
Bacillus halodurans strain C12 ...
Bacillus halodurans, Bacillus halodurans C-125
Appl. Environ. Microbiol.
81
3395-3404
2015
1
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1
1
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1
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11
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684700
Thymark
Mutational analysis of the nuc ...
Escherichia coli
Arch. Biochem. Biophys.
470
20-26
2008
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1
4
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1
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1
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1
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1
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1
4
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1
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1
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2
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1
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2
1
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688399
Helt
Mechanism of dTTP inhibition o ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Mol. Biol.
376
554-569
2008
-
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1
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686667
Johansson
Regulation of dCTP deaminase f ...
Escherichia coli
FEBS J.
274
4188-4198
2007
-
-
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1
3
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1
1
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1
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1
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1
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1
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1
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1
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1
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1
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688732
Zhang
Chloroviruses encode a bifunct ...
Paramecium bursaria Chlorella virus 1
J. Virol.
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7662-7671
2007
1
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1
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3
2
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5
1
2
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4
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1
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1
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1
1
1
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2
2
1
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1
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3
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2
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5
1
2
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1
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1
4
1
1
1
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2
2
1
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669375
Johansson
Structures of dCTP deaminase f ...
Escherichia coli
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280
3051-3059
2005
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1
3
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3
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648212
Huffman
Structural basis for recogniti ...
Methanocaldococcus jannaschii
J. Mol. Biol.
331
885-896
2003
-
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1
1
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1
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1
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652395
Bjornberg
A bifunctional dCTP deaminase- ...
Methanocaldococcus jannaschii
J. Biol. Chem.
278
20667-20672
2003
-
-
1
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2
1
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1
2
1
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4
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1
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4
1
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2
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1
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2
1
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1
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1
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4
1
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209525
Estevenon
An Escherichia coli strain def ...
Escherichia coli
Mol. Gen. Genet.
246
514-518
1995
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1
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209524
Weiss
De novo synthesis of thymidyla ...
Escherichia coli
J. Bacteriol.
176
2194-2199
1994
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2
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1
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209522
Wang
Dcd (dCTP deaminase) gene of E ...
Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium
J. Bacteriol.
174
5647-5653
1992
-
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2
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1
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3
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2
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209523
Speed
Deamination of deoxycytidine n ...
Escherichia coli, Rickettsia prowazekii, Salmonella enterica subsp. enterica serovar Typhimurium
J. Bacteriol.
173
4902-4903
1991
-
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3
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209517
Wang
5-Methyl-dCTP deaminase induce ...
Xanthomonas oryzae
J. Virol.
42
42-48
1982
-
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2
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4
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209518
Neuhard
dCTP Deaminase from Salmonella ...
Salmonella enterica subsp. enterica serovar Typhimurium
Methods Enzymol.
55
418-423
1978
-
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3
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3
1
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1
2
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209519
Beck
Deoxycytidine triphosphate dea ...
Salmonella enterica subsp. enterica serovar Typhimurium
J. Biol. Chem.
250
609-616
1975
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7
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209520
Price
Bacteriophage PBS2-induced deo ...
Bacillus subtilis
J. Virol.
14
1314-1317
1974
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7
1
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1
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209521
Tomita
A novel enzyme, dCTP deaminase ...
Bacillus subtilis
Biochim. Biophys. Acta
179
18-27
1969
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