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EC Tree
The enzyme appears in viruses and cellular organisms
Synonyms
deoxynucleoside monophosphate kinase, deoxyribonucleoside monophosphate kinase, dnmp kinase, deoxycytidylate monophosphate kinase, deoxythymidylate monophosphate kinase, deoxyribonucleoside monophosphokinase, deoxyadenylate monophosphate kinase, deoxyguanylate monophosphate kinase,
more
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deoxyadenylate monophosphate kinase
-
-
deoxycytidylate monophosphate kinase
-
-
deoxyguanylate monophosphate kinase
-
-
deoxynucleoside monophosphate kinase
deoxynucleoside-5'-monophosphate kinase
-
-
-
-
deoxyribonucleoside monophosphate kinase
Tequintavirus T5
-
deoxyribonucleoside monophosphokinase
-
-
-
-
deoxythymidylate monophosphate kinase
-
-
kinase (phosphorylating), deoxynucleoside monophosphate
-
-
-
-
T5 bacteriophage dNMPK
Tequintavirus T5
-
deoxynucleoside monophosphate kinase
-
-
-
-
deoxynucleoside monophosphate kinase
-
-
dNMP kinase
-
-
dNMP kinase
Tequintavirus T5
-
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ATP + a 2'-deoxyribonucleoside 5'-phosphate = ADP + a 2'-deoxyribonucleoside 5'-diphosphate
-
-
-
-
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phospho group transfer
-
-
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-
phospho group transfer
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phospho group transfer
Tequintavirus T5
-
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ATP:deoxynucleoside-phosphate phosphotransferase
dATP can substitute for ATP.
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ATP + 5-bromo-dUMP
ADP + 5-bromo-dUDP
Tequintavirus T5
-
phosphorylated at 45% the rate of dAMP
-
-
r
ATP + deoxynucleoside monophosphate
ADP + deoxynucleoside diphosphate
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
ATP + dUMP
ADP + dUDP
Tequintavirus T5
-
-
-
-
?
additional information
?
-
ATP + dAMP
ADP + dADP
-
-
-
-
r
ATP + dAMP
ADP + dADP
Tequatrovirus T4
-
-
-
-
?
ATP + dAMP
ADP + dADP
Tequintavirus T5
-
-
-
?
ATP + dAMP
ADP + dADP
Tequintavirus T5
-
dAMP is the best acceptor substrate
-
r
ATP + dAMP
ADP + dADP
Tequintavirus T5
-
dAMP is the best acceptor substrate
-
r
ATP + dAMP
ADP + dADP
Tequintavirus T5
-
dATP can replace ATP with less efficiency
-
r
ATP + dAMP
ADP + dADP
Tequintavirus T5
best substrate dAMP, wild-type enzyme
-
-
?
ATP + dCMP
ADP + dCDP
-
-
-
-
r
ATP + dCMP
ADP + dCDP
Tequintavirus T5
-
-
-
?
ATP + dCMP
ADP + dCDP
Tequintavirus T5
-
phosphorylated at 36% the rate of dAMP
-
-
r
ATP + dCMP
ADP + dCDP
Tequintavirus T5
-
phosphorylated at 30% the rate of dAMP, dATP can replace ATP
-
r
ATP + dCMP
ADP + dCDP
Tequintavirus T5
35.2% of the activity with dAMP, wild-type enzyme
-
-
?
ATP + deoxynucleoside monophosphate
ADP + deoxynucleoside diphosphate
-
key step in de novo DNA precursor biosynthesis
-
-
r
ATP + deoxynucleoside monophosphate
ADP + deoxynucleoside diphosphate
Tequintavirus T5
key step in de novo DNA precursor biosynthesis
-
-
?
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
Tequatrovirus T4
-
enzyme induced during viral infection
-
-
?
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
Tequintavirus T5
-
enzyme induced during viral infection
-
r
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
Tequintavirus T5
-
enzyme induced during viral infection
-
r
ATP + dGMP
ADP + dGDP
-
-
-
-
r
ATP + dGMP
ADP + dGDP
Tequintavirus T5
-
-
-
?
ATP + dGMP
ADP + dGDP
Tequintavirus T5
-
dATP can replace ATP
-
r
ATP + dGMP
ADP + dGDP
Tequintavirus T5
-
phosphorylated at 65% the rate of dAMP
-
r
ATP + dGMP
ADP + dGDP
Tequintavirus T5
-
phosphorylated at 65% the rate of dAMP
-
-
r
ATP + dGMP
ADP + dGDP
Tequintavirus T5
49.9% of the activity with dAMP, wild-type enzyme
-
-
?
ATP + dTMP
ADP + dTDP
-
-
-
-
r
ATP + dTMP
ADP + dTDP
Tequintavirus T5
-
-
-
?
ATP + dTMP
ADP + dTDP
Tequintavirus T5
-
dATP can replace ATP with less efficiency
-
r
ATP + dTMP
ADP + dTDP
Tequintavirus T5
-
phosphorylated at 70% the rate of dAMP
-
r
ATP + dTMP
ADP + dTDP
Tequintavirus T5
-
phosphorylated at 70% the rate of dAMP
-
r
ATP + dTMP
ADP + dTDP
Tequintavirus T5
56.4% of the activity with dAMP, wild-type enzyme
-
-
?
additional information
?
-
Tequintavirus T5
-
very poor acceptor substrates are AMP, CMP or dUMP
-
-
?
additional information
?
-
Tequintavirus T5
-
no acceptor substrates are dIMP, deoxyxanthylate, 5-hydroxymethyldeoxycytidylate, UMP, GMP, 5-bromo-dUMP, 5-methyl-dCMP or deoxythymidine
-
-
?
additional information
?
-
Tequintavirus T5
-
no donor substrates are UTP, GTP, CTP, dTTP, dGTP or dCTP
-
-
?
additional information
?
-
Tequintavirus T5
substrate specificities of wild-type and mutant enzymes, structure-function analysis, overview
-
-
?
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ATP + dAMP
ADP + dADP
Tequintavirus T5
-
-
-
?
ATP + dCMP
ADP + dCDP
Tequintavirus T5
-
-
-
?
ATP + deoxynucleoside monophosphate
ADP + deoxynucleoside diphosphate
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
ATP + dGMP
ADP + dGDP
Tequintavirus T5
-
-
-
?
ATP + dTMP
ADP + dTDP
Tequintavirus T5
-
-
-
?
ATP + deoxynucleoside monophosphate
ADP + deoxynucleoside diphosphate
-
key step in de novo DNA precursor biosynthesis
-
-
r
ATP + deoxynucleoside monophosphate
ADP + deoxynucleoside diphosphate
Tequintavirus T5
key step in de novo DNA precursor biosynthesis
-
-
?
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
Tequatrovirus T4
-
enzyme induced during viral infection
-
-
?
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
Tequintavirus T5
-
enzyme induced during viral infection
-
r
ATP + deoxynucleoside phosphate
ADP + deoxynucleoside diphosphate
Tequintavirus T5
-
enzyme induced during viral infection
-
r
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Co2+
Tequintavirus T5
-
requirement, can replace Mg2+
Fe2+
Tequintavirus T5
-
activation, less effective than Mg2+, Co2+, Mn2+
Mn2+
Tequintavirus T5
-
requirement, can replace Co2+ or Mg2+
additional information
Tequintavirus T5
-
no activation by Ba2+, Ca2+, Cd2+, Cr3+, Cu2+, Fe3+, Hg2+, Ni2+, Zn2+
Mg2+
-
dAMP reacts only with ATP after ATP forms the bivalent metal-ion complex MgATP2-, Mg2+ orients and directs ATP and dAMP to the corresponding active sites
Mg2+
-
dCMP reacts only with ATP after ATP forms the bivalent metal-ion complex MgATP2-, Mg2+ orients and directs ATP and dCMP to the corresponding active sites
Mg2+
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dGMP reacts only with ATP after ATP forms the bivalent metal-ion complex MgATP2-, Mg2+ orients and directs ATP and dGMP to the corresponding active sites
Mg2+
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dTMP reacts only with ATP after ATP forms the bivalent metal-ion complex MgATP2-, Mg2+ orients and directs ATP and dTMP to the corresponding active sites
Mg2+
Tequintavirus T5
required
Mg2+
Tequintavirus T5
-
requirement, can replace Co2+
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dAMP
Tequintavirus T5
-
competitive, dTMP as substrate
dCMP
Tequintavirus T5
-
competitive, dTMP as substrate
dGMP
Tequintavirus T5
-
competitive, dTMP as substrate
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Infections
Slow switchover from host RNA synthesis to bacteriophage RNA synthesis after infection of Escherichia coli with a T4 mutant defective in the bacteriophage T4-induced unfolding of the host nucleoid.
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2.8
dUMP
Tequintavirus T5
-
pH 7.5, 37°C
0.11
ATP
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at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.28
ATP
-
at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.36
ATP
Tequintavirus T5
-
pH 7.5, 37°C
0.48
ATP
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at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.67
ATP
-
at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.22
dAMP
Tequintavirus T5
-
pH 7.5, 37°C
1.1
dAMP
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at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.034
dCMP
Tequintavirus T5
-
pH 7.5, 37°C
0.53
dCMP
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at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.22
dGMP
Tequintavirus T5
-
pH 7.5, 37°C
0.33
dGMP
-
at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
0.173
dTMP
Tequintavirus T5
-
pH 7.5, 37°C
0.89
dTMP
-
at 30°C, pH 8.0 and saturating Mg2+ and K+ concentrations
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0.15
dAMP
Tequintavirus T5
-
dTMP as substrate, pH 7.6, 37°C
0.032
dCMP
Tequintavirus T5
-
dTMP as substrate, pH 7.6, 37°C
0.15
dGMP
Tequintavirus T5
-
dTMP as substrate, pH 7.6, 37°C
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14.7
Tequintavirus T5
-
purified enzyme
153.6
Tequintavirus T5
homogeneous recombinant enzyme with dGMP as substrate
25.78
Tequintavirus T5
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dCMP
29
Tequintavirus T5
recombinant protein in sonicated cell exctracts, with dGMP as substrate
36
Tequintavirus T5
recombinant protein after Toyopearl 650 M flow-through, with dGMP as substrate
36.54
Tequintavirus T5
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dGMP
40
Tequintavirus T5
recombinant protein after ammonium sulphate precipitation, with dGMP as substrate
41.24
Tequintavirus T5
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dTMP
73.17
Tequintavirus T5
pH 7.5, 37°C, recombinant wild-type enzyme, substrate dAMP
84.2
Tequintavirus T5
homogeneous recombinant enzyme with dCMP as substrate
additional information
Tequintavirus T5
specific activties of wild-type enzyme and enzyme mutants with different dNMP acceptor substrates, overview
155
Tequintavirus T5
affinity chromatography eluant of the recombinant protein, with dGMP as substrate
155
Tequintavirus T5
ion-exchange chromatography MonoQ eluant of the recombinant protein, with dGMP as substrate
166.8
Tequintavirus T5
-
purified enzyme, substrate dTMP
166.8
Tequintavirus T5
homogeneous recombinant enzyme with dTMP as substrate
232
Tequintavirus T5
-
purified enzyme, substrate dAMP
232
Tequintavirus T5
homogeneous recombinant enzyme with dAMP as substrate
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7 - 7.5
Tequintavirus T5
-
-
7.5 - 8.5
Tequintavirus T5
all enzyme mutants but T17S
7.6
Tequintavirus T5
-
substrate dTMP
additional information
-
between pH 7-8
7
Tequintavirus T5
-
-
7
Tequintavirus T5
wild-type enzyme and mutant T17S
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6.2 - 9
Tequintavirus T5
-
about 80% of maximal activity at pH 6.2 and about 65% of maximal activity at pH 9 with substrate dGMP
6.2 - 9.4
Tequintavirus T5
-
about 65% of maximal activity at pH 6.2 and about 70% of maximal activity at pH 9.4 with substrate dAMP
6.3 - 9
Tequintavirus T5
-
at pH 6.3: about 70% of maximal activity with substrate dTMP, about 85% of maximal activity with substrate dCMP, at pH 9.0: about 50% of maximal activity with substrate dTMP, 65% of maximal activity with substrate dCMP
additional information
Tequintavirus T5
-
active over a broad range
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25
Tequintavirus T5
assay at
37
Tequintavirus T5
-
assay at
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4.2
Tequintavirus T5
-
determined under partially denaturing condiions
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no activity in Escherichia coli
-
-
-
brenda
ATCC 2610
-
-
brenda
Tequatrovirus T4
from infected Escherichia coli cells
-
-
brenda
Tequatrovirus T4
maturation defective phage mutant T4amBL292, phage-coded activity which is a component of T4 dNTP-synthezising enzyme complex
-
-
brenda
Tequintavirus T5
-
SwissProt
brenda
Tequintavirus T5
from infected Escherichia coli cells
-
-
brenda
Tequintavirus T5
gene dnk
SwissProt
brenda
Tequintavirus T5
mutant T5amH128a
-
-
brenda
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additional information
Tequintavirus T5
essential catalytic site residue, active site modelling, overview. The charged amino acid residues of the NMP binding domain and the presence of an OH-group at position 17 are important for the catalytic activity. Arginine residues at positions 130 and 172 are involved in the binding to the donor gamma-phosphoryl and acceptor alpha-phosphoryl groups, as well as the aspartic acid residue at position 16 of the ATP-binding site (P-loop), in the binding to some acceptors, first of all dTMP. The NMPK-P loop, or a glycine enriched loop, forms an anion hole interacting with donor phosphoryl groups. It contains a GX1X2X3X4GKX5T(S) consensus called the Walker A motif
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20640
-
calculated from amino acid sequence
22000
Tequatrovirus T4
-
-
22930
-
calculated from amino acid sequence
24260
-
calculated from amino acid sequence
24690
-
calculated from amino acid sequence
28670
Tequintavirus T5
calculated from nucleotid sequence
29000
Tequintavirus T5
-
1 * 29000, SDS-PAGE
29140
Tequintavirus T5
SDS-PAGE
29140
Tequintavirus T5
-
analytical equilibrium ultracentrifugation
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monomer
Tequintavirus T5
-
monomer
Tequintavirus T5
-
1 * 29000, SDS-PAGE
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D16N
Tequintavirus T5
site-directed mutagenesis of the P loop-related residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
D170N
Tequintavirus T5
site-directed mutagenesis of the core residue, almost inactive mutant
E176Q
Tequintavirus T5
site-directed mutagenesis of the core residue, almost inactive mutant
G137A
Tequintavirus T5
site-directed mutagenesis of the dNMP-binding residue, the mutant shows highly reduced activity and altered substrate specificity compared to the wild-type enzyme
K131E
Tequintavirus T5
site-directed mutagenesis, of the dNMP-binding residue, the mutant shows slightly increased activity and altered substrate specificity compared to the wild-type enzyme
Q134A
Tequintavirus T5
site-directed mutagenesis of the dNMP-binding residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
R130K
Tequintavirus T5
site-directed mutagenesis of the dNMP-binding residue, almost inactive mutant
R172I
Tequintavirus T5
site-directed mutagenesis of the core residue, almost inactive mutant
S13A
Tequintavirus T5
site-directed mutagenesis of the P loop-related residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
T138A
Tequintavirus T5
site-directed mutagenesis of the dNMP-binding residue, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
T17N
Tequintavirus T5
site-directed mutagenesis of the P loop-related residue, the mutant shows highly reduced activity and altered substrate specificity compared to the wild-type enzyme
T17S
Tequintavirus T5
site-directed mutagenesis, of the P loop-related residue, the mutant shows reduced activity compared to the wild-type enzyme
W150A
Tequintavirus T5
site-directed mutagenesis of the core residue
W150F
Tequintavirus T5
site-directed mutagenesis of the conserved residue, the mutant shows unaltered activity compared to the wild-type enzyme
additional information
Tequintavirus T5
all enzyme mutants but T17S show an increased pH optimum compared to the wild-type enzyme
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37
Tequintavirus T5
-
60 min, stable
42
Tequintavirus T5
-
10% loss of activity after 30 min
47
Tequintavirus T5
-
50% loss of activity after 30 min
52
Tequintavirus T5
-
rapid inactivation, t1/2: 5 min
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-20°C, purified enzyme, 15% glycerol, only loss of a small part of activity after 6 months
Tequintavirus T5
-
0°C, 25% loss of activity per day in crude cell extract
Tequintavirus T5
-
4°C, several months, ionic strength 0.1 or above
Tequintavirus T5
-
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1190fold to homogeneity
Tequintavirus T5
-
500fold
Tequintavirus T5
-
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange and affinity chromatography
Tequintavirus T5
two alternative methods, the first involves ion-exchange chromatography on Mono Q, the second involves affinity chromatography on Toyopearl AF-Red, the protein is purified 5.34fold in both cases
Tequintavirus T5
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expression in Escherichia coli
Tequintavirus T5
expression in Escherichia coli strain BL21 (DE3)
-
expression in Escherichia coli strain BL21 (DE3), CK expression at 37°C forms insoluble inclusion bodoes, therefore expression was performed at 25°C
-
gene dnk, recombinant expression of wild-type and mutant enzymes in Escherichia coli strains BL21(DE3) and XL10Gold
Tequintavirus T5
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Allen, J.R.; Lasser, G.W.; Goldman, D.A.; Booth, J.W.; Mathews, C.K.
T4 phage deoxyribonucleotide-synthesizing enzyme complex. Further studies on enzyme composition and regulation
J. Biol. Chem.
258
5746-5753
1983
Tequatrovirus T4, no activity in Escherichia coli
brenda
Bessman, M.J.; Herriott, S.T.; Van Bibber Orr, M.J.
The enzymology of virus-infected bacteria. VI. Purification and properties of the deoxynucleotide kinase induced by bacteriophage T5
J. Biol. Chem.
240
439-445
1965
Tequintavirus T5, no activity in Escherichia coli
brenda
Mikoulinskaia, G.V.; Gubanov, S.I.; Zimin, A.A.; Kolesnikov, I.V.; Feofanov, S.A.; Miroshnikov, A.I.
Purification and characterization of the deoxynucleoside monophosphate kinase of bacteriophage T5
Protein Expr. Purif.
27
195-201
2003
Tequintavirus T5, no activity in Escherichia coli
brenda
Bao, J.; Ryu, D.D.
Cloning of deoxynucleoside monophosphate kinase genes and biosynthesis of deoxynucleoside diphosphates
Biotechnol. Bioeng.
93
572-580
2006
Saccharomyces cerevisiae
brenda
Mikoulinskaia, G.V.; Zimin, A.A.; Feofanov, S.A.; Miroshnikov, A.I.
Identification, cloning, and expression of bacteriophage T5 dnk gene encoding a broad specificity deoxyribonucleoside monophosphate kinase (EC 2.7.4.13)
Protein Expr. Purif.
33
166-175
2004
Tequintavirus T5 (Q6QGP4)
brenda
Mikoulinskaia, G.; Taran, S.; Skoblov, Y.; Feofanov, S.
The study of the bacteriophage T5 deoxynucleoside monophosphate kinase active site by site-directed mutagenesis
Russ. J. Bioorg. Chem.
39
607-618
2013
Tequintavirus T5 (Q6QGP4)
-
brenda
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