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Information on EC 3.6.1.65 - (d)CTP diphosphatase for references in articles please use BRENDA:EC3.6.1.65
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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 expected taxonomic range for this enzyme is: Escherichia coli
Synonyms
(d)CTP diphosphohydrolase, (d)CTP pyrophosphohydrolase, 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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gene name
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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
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
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3% of the activity with 5-methyl-dCTP
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-
?
CDP + H2O
cytidine + diphosphate
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2% of the activity with 5-methyl-dCTP
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-
?
CTP + H2O
CMP + diphosphate
dCTP + H2O
dCMP + diphosphate
dTTP + H2O
dTMP + diphosphate
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2% of the activity with 5-methyl-dCTP
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-
?
dUTP + H2O
dUMP + diphosphate
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1% of the activity with 5-methyl-dCTP
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?
additional information
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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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-
-
-
?
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
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-
-
-
?
CTP + H2O
CMP + diphosphate
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-
-
?
CTP + H2O
CMP + diphosphate
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48% of the activity with 5-methyl-dCTP
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?
dCTP + H2O
dCMP + diphosphate
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-
-
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?
dCTP + H2O
dCMP + diphosphate
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-
-
?
dCTP + H2O
dCMP + diphosphate
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23% of the activity with 5-methyl-dCTP
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?
dCTP + H2O
dCMP + 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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Mn2+
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about 10% of the activity with Mg2+
additional information
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absolute requirement for a divalnt cation. No activity in presence of Zn2+, Co2+, Ca2+
Mg2+
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required, activity constant 1.2 mM
Mg2+
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most effective cation, with optimal activity at 5 mM Mg2+
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0.0055 - 0.31
2-hydroxy-dATP
0.0557
5-hydroxy-CTP
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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
0.0055
2-hydroxy-dATP
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mutant D118E, 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
0.31
2-hydroxy-dATP
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mutant R72A, 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
0.0069
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
0.028
5-Methyl-dCTP
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wild-type, 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
0.22
5-Methyl-dCTP
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mutant E33A, 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
0.428
CTP
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pH 8.5, 30°C
0.65
dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
0.767
dCTP
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pH 9.0, 37°C
0.77
dCTP
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mutant D118N, 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
0.99
dCTP
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wild-type, 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
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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
0.17
2-hydroxy-dATP
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wild-type, pH not specified in the publication, 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
2.7
2-hydroxy-dATP
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mutant D118A, 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
6.5
5-Methyl-dCTP
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mutant D118E, 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
9.1
5-Methyl-dCTP
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mutant D118A, 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
35
5-Methyl-dCTP
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wild-type, pH not specified in the publication, temperature not specified in the publication
51.7
5-Methyl-dCTP
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pH 9.0, 37°C
9.28
CTP
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pH 8.5, 30°C
2.1
dCTP
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mutant D118E, 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
6.3
dCTP
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wild-type, pH not specified in the publication, 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
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630
5-hydroxy-CTP
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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
4.6
2-hydroxy-dATP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
6.4
2-hydroxy-dATP
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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
21
2-hydroxy-dATP
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mutant E33A, 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
13
5-Methyl-dCTP
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mutant E33A, 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
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pH 9.0, 37°C
780
5-Methyl-dCTP
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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
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wild-type, 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
21.7
CTP
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pH 8.5, 30°C
2.3
dCTP
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mutant D118E, pH not specified in the publication, temperature not specified in the publication
3.5
dCTP
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mutant D118N, 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
14
dCTP
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mutant R72A, pH not specified in the publication, temperature not specified in the publication
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193
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substrate 5-methyl-dCTP, pH 9.0, 37°C
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brenda
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brenda
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UniProt
brenda
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D118A
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impaired hydrolysis of substrate 2-hydroxy-dATP
D118E
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increase in hydrolysis of substrate 2-hydroxy-dATP
D118N
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impaired hydrolysis of substrate 2-hydroxy-dATP
E33A
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increase in hydrolysis of substrate 2-hydroxy-dATP
E33Q
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increase in hydrolysis of substrate 2-hydroxy-dATP
E43A
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substrate 5-methyl-dCTP, about 20% of wild-type activity, substrate dCTP, 1%of wild-type activity
E52A
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complete loss of activity
E52D
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complete loss of activity
E52Q
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complete loss of activity
E55A
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substrate 5-methyl-dCTP, about 20% of wild-type activity, substrate dCTP, 1%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
E55Q
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substrate 5-methyl-dCTP, about 110% of wild-type activity, substrate dCTP, 10% of wild-type activity
E56A
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complete loss of activity
E56D
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complete loss of activity
E56Q
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complete loss of activity
G36A
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complete loss of activity
G37A
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complete loss of activity
K38A
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substrate 5-methyl-dCTP, about 30% of wild-type activity, substrate dCTP, 10% of wild-type activity
K38R
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substrate 5-methyl-dCTP, about 40% of wild-type activity, substrate dCTP, 5% 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
R51A
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substrate 5-methyl-dCTP, about 5% of wild-type activity, substrate dCTP, 1% of wild-type activity
R72A
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impaired hydrolysis of substrate 2-hydroxy-dATP
D118A
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impaired 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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E33A
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increase in 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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R72A
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impaired hydrolysis of substrate 2-hydroxy-dATP
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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
brenda
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
brenda
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, Escherichia coli (P77788)
brenda
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, Escherichia coli (P77788)
brenda
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3.6.1.65 (d)CTP diphosphatase
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