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EC Tree
IUBMB Comments The enzyme, characterized from the bacterium Escherichia coli, is specific for the pyrimidine nucleotides CTP and dCTP. It also acts on 5-methyl-dCTP, 5-hydroxy-dCTP and 8-hydroxy-dGTP.
The enzyme appears in viruses and cellular organisms
Synonyms
(d)CTP diphosphohydrolase, (d)CTP pyrophosphohydrolase, MazG,
nudG , Orf135,
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(d)CTP diphosphohydrolase
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(d)CTP pyrophosphohydrolase
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nudG
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-
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CTP + H2O = CMP + diphosphate
(1)
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-
dCTP + H2O = dCMP + diphosphate
(2)
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-
-
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(deoxy)cytidine 5'-triphosphate diphosphohydrolase
The enzyme, characterized from the bacterium Escherichia coli, is specific for the pyrimidine nucleotides CTP and dCTP. It also acts on 5-methyl-dCTP, 5-hydroxy-dCTP and 8-hydroxy-dGTP.
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2-hydroxy-dATP + H2O
2-hydroxy-AMP + diphosphate
2-hydroxy-dATP + H2O
2-hydroxy-dAMP + diphosphate
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-
-
-
?
5-hydroxy-CTP + H2O
5-hydroxy-CMP + diphosphate
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-
-
?
5-hydroxy-dCTP + H2O
5-hydroxy-dCMP + diphosphate
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4.3% of the activity with 5-hydroxy-CTP
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?
5-methyl-dCTP + H2O
5-methyl-dCMP + diphosphate
8-hydroxy-dGTP + H2O
8-hydroxy-dGMP + diphosphate
arabinosyl-CTP + H2O
arabinosyl-CMP + diphosphate
3% of the activity with 5-methyl-dCTP
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-
?
ATP + H2O
AMP + diphosphate
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-
-
-
?
CDP + H2O
cytidine + diphosphate
2% of the activity with 5-methyl-dCTP
-
-
?
CTP + H2O
CMP + diphosphate
dATP + H2O
dAMP + diphosphate
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-
-
-
?
dCTP + H2O
dCMP + diphosphate
dGTP + H2O
dGMP + diphosphate
-
preferred substrate
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-
?
dTTP + H2O
dTMP + diphosphate
dUTP + H2O
dUMP + diphosphate
GTP + H2O
GMP + diphosphate
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-
-
-
?
UTP + H2O
UMP + diphosphate
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-
-
-
?
additional information
?
-
2-hydroxy-dATP + H2O
2-hydroxy-AMP + diphosphate
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-
-
-
?
2-hydroxy-dATP + H2O
2-hydroxy-AMP + diphosphate
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-
-
-
?
5-methyl-dCTP + H2O
5-methyl-dCMP + diphosphate
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-
-
-
?
5-methyl-dCTP + H2O
5-methyl-dCMP + diphosphate
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-
-
?
5-methyl-dCTP + H2O
5-methyl-dCMP + diphosphate
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-
-
-
?
8-hydroxy-dGTP + H2O
8-hydroxy-dGMP + diphosphate
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-
-
-
?
8-hydroxy-dGTP + H2O
8-hydroxy-dGMP + diphosphate
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-
-
-
?
CTP + H2O
CMP + diphosphate
48% of the activity with 5-methyl-dCTP
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-
?
CTP + H2O
CMP + diphosphate
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-
-
?
CTP + H2O
CMP + diphosphate
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-
-
-
?
CTP + H2O
CMP + diphosphate
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-
-
-
?
dCTP + H2O
dCMP + diphosphate
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-
-
-
?
dCTP + H2O
dCMP + diphosphate
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-
-
?
dCTP + H2O
dCMP + diphosphate
23% of the activity with 5-methyl-dCTP
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-
?
dCTP + H2O
dCMP + diphosphate
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-
-
-
?
dCTP + H2O
dCMP + diphosphate
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preferred substrate
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-
?
dTTP + H2O
dTMP + diphosphate
2% of the activity with 5-methyl-dCTP
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-
?
dTTP + H2O
dTMP + diphosphate
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-
-
-
?
dUTP + H2O
dUMP + diphosphate
1% of the activity with 5-methyl-dCTP
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-
?
dUTP + H2O
dUMP + diphosphate
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-
-
-
?
additional information
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enzyme is highly specific for pyrimidine (deoxy)nucleoside triphosphates. Like other Nudix hydrolases, the enzyme cleaves its substrates to produce a nucleoside monophosphate and diphosphate
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additional information
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enzyme is highly specific for pyrimidine (deoxy)nucleoside triphosphates. Like other Nudix hydrolases, the enzyme cleaves its substrates to produce a nucleoside monophosphate and diphosphate
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additional information
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no substrates: CMP, dCMP, dATP, ATP, dGTP, GTP, dUTP, CDP-ethanolamine, CDP-choline, CDP-glucose, CDP-glycerol
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?
additional information
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no substrates: CMP, dCMP, dATP, ATP, dGTP, GTP, dUTP, CDP-ethanolamine, CDP-choline, CDP-glucose, CDP-glycerol
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additional information
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the enzyme shows no binding or hydrolysis activity against (p)ppGpp and 8-oxo-dGTP
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Mn2+
about 10% of the activity with Mg2+
Mg2+
most effective cation, with optimal activity at 5 mM Mg2+
Mg2+
required, activity constant 1.2 mM
additional information
absolute requirement for a divalnt cation. No activity in presence of Zn2+, Co2+, Ca2+
additional information
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absolute requirement for a divalnt cation. No activity in presence of Zn2+, Co2+, Ca2+
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0.0055 - 4.86
2-hydroxy-dATP
0.0557
5-hydroxy-CTP
pH 8.5, 30°C
0.0057 - 0.24
5-Methyl-dCTP
0.41
8-hydroxy-dGTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
1.43
ATP
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pH and temperature not specified in the publication
1.2
dATP
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pH and temperature not specified in the publication
0.14
dGTP
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pH and temperature not specified in the publication
1.23
dTTP
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pH and temperature not specified in the publication
3.24
dUTP
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pH and temperature not specified in the publication
0.6
UTP
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pH and temperature not specified in the publication
0.0055
2-hydroxy-dATP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
4.86
2-hydroxy-dATP
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pH and temperature not specified in the publication
0.31
2-hydroxy-dATP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
0.027
2-hydroxy-dATP
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wild-type, pH not specified in the publication, temperature not specified in the publication
0.067
2-hydroxy-dATP
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mutant E33A, pH not specified in the publication, temperature not specified in the publication
0.19
2-hydroxy-dATP
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mutant D118A, pH not specified in the publication, temperature not specified in the publication
0.0057
5-Methyl-dCTP
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mutant D118N, pH not specified in the publication, temperature not specified in the publication
0.24
5-Methyl-dCTP
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mutant D118A, pH not specified in the publication, temperature not specified in the publication
0.22
5-Methyl-dCTP
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mutant E33A, pH not specified in the publication, temperature not specified in the publication
0.172
5-Methyl-dCTP
pH 9.0, 37°C
0.0069
5-Methyl-dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
0.015
5-Methyl-dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
0.172
5-Methyl-dCTP
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pH 9.0, 37°C
0.028
5-Methyl-dCTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
0.428
CTP
pH 8.5, 30°C
0.85
CTP
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pH and temperature not specified in the publication
0.77
dCTP
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mutant D118N, pH not specified in the publication, temperature not specified in the publication
1.16
dCTP
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pH and temperature not specified in the publication
0.99
dCTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
0.93
dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
0.65
dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
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0.17 - 2.7
2-hydroxy-dATP
35.1
5-hydroxy-CTP
pH 8.5, 30°C
0.89
8-hydroxy-dGTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
1.48
ATP
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pH and temperature not specified in the publication
1.05
dATP
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pH and temperature not specified in the publication
6.67
dGTP
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pH and temperature not specified in the publication
3.68
dTTP
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pH and temperature not specified in the publication
4.94
dUTP
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pH and temperature not specified in the publication
0.19
GTP
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pH and temperature not specified in the publication
2.15
UTP
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pH and temperature not specified in the publication
0.45
2-hydroxy-dATP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
1.4
2-hydroxy-dATP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
1.42
2-hydroxy-dATP
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mutant E33A, pH not specified in the publication, temperature not specified in the publication
1.93
2-hydroxy-dATP
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pH and temperature not specified in the publication
2.7
2-hydroxy-dATP
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mutant D118A, pH not specified in the publication, temperature not specified in the publication
0.17
2-hydroxy-dATP
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wild-type, pH not specified in the publication, temperature not specified in the publication
6.5
5-Methyl-dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
51.7
5-Methyl-dCTP
pH 9.0, 37°C
35
5-Methyl-dCTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
11
5-Methyl-dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
9.1
5-Methyl-dCTP
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mutant D118A, pH not specified in the publication, temperature not specified in the publication
7.7
5-Methyl-dCTP
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mutant D118N, pH not specified in the publication, temperature not specified in the publication
2.8
5-Methyl-dCTP
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mutant E33A, pH not specified in the publication, temperature not specified in the publication
1.62
CTP
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pH and temperature not specified in the publication
8.8
dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
2.7
dCTP
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mutant D118N, pH not specified in the publication, temperature not specified in the publication
2.1
dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
6.3
dCTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
5.75
dCTP
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pH and temperature not specified in the publication
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630
5-hydroxy-CTP
pH 8.5, 30°C
2.2
8-hydroxy-dGTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
82
2-hydroxy-dATP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
4.6
2-hydroxy-dATP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
21
2-hydroxy-dATP
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mutant E33A, pH not specified in the publication, temperature not specified in the publication
6.4
2-hydroxy-dATP
-
wild-type, pH not specified in the publication, temperature not specified in the publication
14
2-hydroxy-dATP
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mutant D118A, pH not specified in the publication, temperature not specified in the publication
1400
5-Methyl-dCTP
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mutant D118N, pH not specified in the publication, temperature not specified in the publication
38
5-Methyl-dCTP
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mutant D118A, pH not specified in the publication, temperature not specified in the publication
301
5-Methyl-dCTP
pH 9.0, 37°C
13
5-Methyl-dCTP
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mutant E33A, pH not specified in the publication, temperature not specified in the publication
780
5-Methyl-dCTP
-
mutant R72A, pH not specified in the publication, temperature not specified in the publication
944
5-Methyl-dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
1300
5-Methyl-dCTP
-
wild-type, pH not specified in the publication, temperature not specified in the publication
21.7
CTP
pH 8.5, 30°C
18
dCTP
pH 9.0, 37°C
14
dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
6.4
dCTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
3.5
dCTP
-
mutant D118N, pH not specified in the publication, temperature not specified in the publication
2.3
dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
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193
substrate 5-methyl-dCTP, pH 9.0, 37°C
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UniProt
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E52D
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complete loss of activity
K38R
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substrate 5-methyl-dCTP, about 40% of wild-type activity, substrate dCTP, 5% of wild-type activity
K38A
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substrate 5-methyl-dCTP, about 30% of wild-type activity, substrate dCTP, 10% of wild-type activity
G37A
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complete loss of activity
G36A
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complete loss of activity
E56Q
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complete loss of activity
E56D
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complete loss of activity
E56A
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complete loss of activity
E55Q
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substrate 5-methyl-dCTP, about 110% of wild-type activity, substrate dCTP, 10% of wild-type activity
E55D
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substrate 5-methyl-dCTP, about 40% of wild-type activity, substrate dCTP, 2% of wild-type activity
E55A
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substrate 5-methyl-dCTP, about 20% of wild-type activity, substrate dCTP, 1%of wild-type activity
E52Q
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complete loss of activity
E52A
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complete loss of activity
E43A
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substrate 5-methyl-dCTP, about 20% of wild-type activity, substrate dCTP, 1%of wild-type activity
L53A
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substrate 5-methyl-dCTP, about 30% of wild-type activity, substrate dCTP, 30% of wild-type activity
E33Q
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increase in hydrolysis of substrate 2-hydroxy-dATP
R51A
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substrate 5-methyl-dCTP, about 5% of wild-type activity, substrate dCTP, 1% of wild-type activity
E33A
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increase in hydrolysis of substrate 2-hydroxy-dATP
R72A
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impaired hydrolysis of substrate 2-hydroxy-dATP
D118N
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impaired hydrolysis of substrate 2-hydroxy-dATP
D118E
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increase in hydrolysis of substrate 2-hydroxy-dATP
D118A
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impaired hydrolysis of substrate 2-hydroxy-dATP
R72A
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impaired hydrolysis of substrate 2-hydroxy-dATP
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E33Q
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increase in hydrolysis of substrate 2-hydroxy-dATP
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E33A
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increase in hydrolysis of substrate 2-hydroxy-dATP
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D118N
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impaired hydrolysis of substrate 2-hydroxy-dATP
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D118A
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impaired hydrolysis of substrate 2-hydroxy-dATP
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expressed in Escherichia coli BL21(DE3) cells
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expression as glutathione S-transferase fusion protein
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Kamiya, H.; Iida, E.; Harashima, H.
Important amino acids in the phosphohydrolase module of Escherichia coli Orf135
Biochem. Biophys. Res. Commun.
323
1063-1068
2004
Escherichia coli
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Iida, E.; Satou, K.; Mishima, M.; Kojima, C.; Harashima, H.; Kamiya, H.
Amino acid residues involved in substrate recognition of the Escherichia coli Orf135 protein
Biochemistry
44
5683-5689
2005
Escherichia coli, Escherichia coli JD22899
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Fujikawa, K.; Kasai, H.
The oxidized pyrimidine ribonucleotide, 5-hydroxy-CTP, is hydrolyzed efficiently by the Escherichia coli recombinant Orf135 protein
DNA Repair
1
571-576
2002
Escherichia coli (P77788), Escherichia coli
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OHandley, S.; Dunn, C.; Bessman, M.
Orf135 from Escherichia coli is a nudix hydrolase specific for CTP, dCTP, and 5-methyl-dCTP
J. Biol. Chem.
276
5421-5426
2001
Escherichia coli (P77788), Escherichia coli
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Rihtman, B.; Bowman-Grahl, S.; Millard, A.; Corrigan, R.M.; Clokie, M.R.J.; Scanlan, D.J.
Cyanophage MazG is a pyrophosphohydrolase but unable to hydrolyse magic spot nucleotides
Environ. Microbiol. Rep.
11
448-455
2019
Synechococcus phage S-PM2
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