Information on EC 2.7.1.76 - 2'-deoxyadenosine kinase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
2.7.1.76
-
RECOMMENDED NAME
GeneOntology No.
2'-deoxyadenosine kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + 2'-deoxyadenosine = ADP + dAMP
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
purine deoxyribonucleosides salvage
-
-
purine metabolism
-
-
Purine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:2'-deoxyadenosine 5'-phosphotransferase
2'-Deoxyguanosine can also act as acceptor. Possibly identical with EC 2.7.1.74 deoxycytidine kinase.
CAS REGISTRY NUMBER
COMMENTARY hide
37278-12-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
TrEMBL
Manually annotated by BRENDA team
gene yaaF, yaaF encodes an enzyme with specificity for deoxyadenosine and deoxycytidine
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-deoxyadenosine-3'-fluoro-3'-triphosphate + 2'-deoxyadenosine
2'-deoxyadenosine-3'-fluoro-3'-diphosphate + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
-
-
-
-
?
2'-deoxyadenosine-3'-fluoro-3'-triphosphate + 2'-deoxycytidine
2'-deoxyadenosine-3'-fluoro-3'-diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
2'-deoxyadenosine-3'-fluoroara-3'-triphosphate + 2'-deoxyadenosine
2'-deoxyadenosine-3'-fluoroara-3'-diphosphate + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
-
low activity
-
-
?
2'-deoxyadenosine-3'-fluoroara-3'-triphosphate + 2'-deoxycytidine
2'-deoxyadenosine-3'-fluoroara-3'-diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
low activity
-
-
?
2'-deoxyadenosine-3'-triphosphate + 2'-deoxyadenosine
2'-deoxyadenosine-3'-diphosphate + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
-
-
-
-
?
2'-deoxyadenosine-3'-triphosphate + 2'-deoxycytidine
2'-deoxyadenosine-3'-diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
3'-deoxyadenosine 2'-triphosphate + 2'-deoxyadenosine
3'-deoxyadenosine-2'-diphosphate + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
-
-
-
-
?
3'-deoxyadenosine 2'-triphosphate + 2'-deoxycytidine
3'-deoxyadenosine-2'-diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
3'-deoxyadenosine-3'-fluoro-2'-triphosphate + 2'-deoxyadenosine
3'-deoxyadenosine-3'-fluoro-2'-diphosphate + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
-
-
-
-
?
3'-deoxyadenosine-3'-fluoro-2'-triphosphate + 2'-deoxycytidine
3'-deoxyadenosine-3'-fluoro-2'-diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
3'-deoxyadenosine-3'-fluoroxylo-2'-triphosphate + 2'-deoxyadenosine
3'-deoxyadenosine-3'-fluoroxylo-2'-diphosphate + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
-
-
-
-
?
3'-deoxyadenosine-3'-fluoroxylo-2'-triphosphate + 2'-deoxycytidine
3'-deoxyadenosine-3'-fluoroxylo-2'-diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
ATP + 1-beta-D-arabinofuranosyl cytosine
ADP + 1-beta-D-arabinofuranosyl cytosine 5'-phosphate
show the reaction diagram
ATP + 1-beta-D-arabinofuranosyl uracil
ADP + 1-beta-D-arabinofuranosyl uracil 5'-phosphate
show the reaction diagram
0.3% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide
ADP + 1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide 5'-monophosphate
show the reaction diagram
-
trivial name ribavirin
-
?
ATP + 2',2'-difluoro-2'-deoxycytidine
ADP + 2',2'-difluoro-2'-deoxycytidine 5'-phosphate
show the reaction diagram
20% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2',2'-difluoro-2'-deoxycytidine
ADP + 2',2'-difluoro-2'-deoxycytidine monophosphate
show the reaction diagram
20% relative activity compared to deoxyadenosine
-
-
?
ATP + 2',2'-difluoro-2'-deoxyguanosine
ADP + 2',2'-difluoro-2'-deoxyguanosine 5'-phosphate
show the reaction diagram
24% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2',2'-difluoro-2'-deoxyguanosine
ADP + 2',2'-difluoro-2'-deoxyguanosine monophosphate
show the reaction diagram
24% relative activity compared to deoxyadenosine
-
-
?
ATP + 2',2'-difluorodeoxycytidine
ADP + 2',2'-difluorodeoxycytidine 5'-phosphate
show the reaction diagram
ATP + 2',3'-dideoxyadenosine
ADP + 2',3'-dideoxyadenosine 5'-monophosphate
show the reaction diagram
1.5% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2',3'-dideoxyadenosine
ADP + 2',3'-dideoxyadenosine monophosphate
show the reaction diagram
1.5% relative activity compared to deoxyadenosine
-
-
?
ATP + 2',3'-dideoxycytidine
ADP + 2',3'-dideoxycytidine 5'-monophosphate
show the reaction diagram
0.1% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2',3'-dideoxyguanosine
ADP + 2',3'-dideoxyguanosine 5'-monophosphate
show the reaction diagram
0.5% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2',3'-dideoxyinoosine
ADP + 2',3'-dideoxyinosine 5'-monophosphate
show the reaction diagram
3.9% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2',3'-dideoxyinosine
ADP + 2',3'-dideoxyinosine 5'-phosphate
show the reaction diagram
3.9% relative activity compared to deoxyadenosine
-
-
?
ATP + 2'-deoxyadenosine
ADP + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
ATP + 2'-deoxycytidine
ADP + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
ATP + 2'-deoxyguanosine
ADP + 2'-deoxyguanosine 5'-phosphate
show the reaction diagram
97% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2'-deoxyinosine
ADP + 2'-deoxyinosine 5'-phosphate
show the reaction diagram
116% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2'-deoxythymidine
ADP + 2'-deoxythymidine 5'-phosphate
show the reaction diagram
0.01% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2'-deoxyuridine
ADP + 2'-deoxyuridine 5'-phosphate
show the reaction diagram
0.9% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2'-fluoro-arabinosyl adenine
ADP + 2'-fluoro-arabinosyl adenine 5'-monophosphate
show the reaction diagram
15% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2'-fluoro-arabinosyl adenine
ADP + 2'-fluoro-arabinosyl adenine monophosphate
show the reaction diagram
ATP + 2-chloro-2'-deoxyadenosine
ADP + 2-chloro-2'-deoxyadenosine 5'-phosphate
show the reaction diagram
111% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 2-chloro-2'-deoxyadenosine
ADP + 2-chloro-2'-deoxyadenosine monophosphate
show the reaction diagram
111% relative activity compared to deoxyadenosine
-
-
?
ATP + 9-beta-D-arabinofuranosyl adenine
ADP + 9-beta-D-arabinofuranosyl adenine 5'-monophosphate
show the reaction diagram
77% relative activity compared to deoxyadenosine
-
-
?
ATP + 9-beta-D-arabinofuranosyl adenine
ADP + 9-beta-D-arabinofuranosyl adenine 5'-phosphate
show the reaction diagram
77% of the activity with 2'-deoxyadenosine
-
-
?
ATP + 9-beta-D-arabinofuranosyl guanine
ADP + 9-beta-D-arabinofuranosyl guanine 5'-monophosphate
show the reaction diagram
6% relative activity compared to deoxyadenosine
-
-
?
ATP + 9-beta-D-arabinofuranosyl guanine
ADP + 9-beta-D-arabinofuranosyl guanine 5'-phosphate
show the reaction diagram
6% of the activity with 2'-deoxyadenosine
-
-
?
ATP + adenosine
ADP + adenosine 5'-monophosphate
show the reaction diagram
7.4% of the activity with 2'-deoxyadenosine
-
-
?
ATP + adenosine
ADP + AMP
show the reaction diagram
ATP + cytidine
ADP + CMP
show the reaction diagram
-
approx. 18% of activity with deoxyadenosine
-
?
ATP + cytidine
ADP + cytidine 5'-monophosphate
show the reaction diagram
0.1% of the activity with 2'-deoxyadenosine
-
-
?
ATP + deoxyadenosine
ADP + 5'-dAMP
show the reaction diagram
ATP + deoxyadenosine
ADP + dAMP
show the reaction diagram
ATP + deoxycytidine
ADP + dCMP
show the reaction diagram
ATP + deoxycytosine
ADP + dCMP
show the reaction diagram
81% relative activity compared to deoxyadenosine
-
-
?
ATP + deoxyguanosine
ADP + dGMP
show the reaction diagram
ATP + deoxyinosine
ADP + dIMP
show the reaction diagram
116% relative activity compared to deoxyadenosine
-
-
?
ATP + deoxyuridine
ADP + dUMP
show the reaction diagram
0.9% relative activity compared to deoxyadenosine
-
-
?
ATP + fludarabine
ADP + fludarabine monophosphate
show the reaction diagram
-
-
-
?
ATP + gemcitabine
ADP + ?
show the reaction diagram
-
-
-
?
ATP + guanosine
ADP + GMP
show the reaction diagram
5.5% relative activity compared to deoxyadenosine
-
-
?
ATP + guanosine
ADP + guanosine 5'-monophosphate
show the reaction diagram
5.5% of the activity with 2'-deoxyadenosine
-
-
?
ATP + inosine
ADP + IMP
show the reaction diagram
5.9% relative activity compared to deoxyadenosine
-
-
?
ATP + inosine
ADP + inosine monophosphate
show the reaction diagram
5.9% of the activity with 2'-deoxyadenosine
-
-
?
ATP + penciclovir
ADP + ?
show the reaction diagram
1.4% relative activity compared to deoxyadenosine
-
-
?
ATP + thymidine
ADP + thymidine 5'-phosphate
show the reaction diagram
-
-
-
?
CTP + 2'-deoxyadenosine
CDP + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
CTP, 3% of the activity with ATP
-
-
?
CTP + deoxyadenosine
CDP + dAMP
show the reaction diagram
dATP + deoxyadenosine
dADP + dAMP
show the reaction diagram
2% activity compared to ATP
-
-
?
dCTP + deoxyadenosine
dCDP + dAMP
show the reaction diagram
106% activity compared to ATP
-
-
?
dTTP + deoxyadenosine
dTDP + dAMP
show the reaction diagram
GTP + 2'-deoxyadenosine
GDP + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
GTP + 2'-deoxycytidine
GDP + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
GTP + deoxyadenosine
GDP + dAMP
show the reaction diagram
triphosphate + 2'-deoxycytidine
diphosphate + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
-
-
-
-
?
TTP + deoxyadenosine
TDP + dAMP
show the reaction diagram
UTP + 2'-deoxyadenosine
UDP + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
UTP, 2% of the activity with ATP
-
-
?
UTP + deoxyadenosine
UDP + dAMP
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + 2'-deoxyadenosine
ADP + 2'-deoxyadenosine 5'-phosphate
show the reaction diagram
ATP + 2'-deoxycytidine
ADP + 2'-deoxycytidine 5'-phosphate
show the reaction diagram
ATP + deoxyadenosine
ADP + 5'-dAMP
show the reaction diagram
ATP + deoxyadenosine
ADP + dAMP
show the reaction diagram
Q54YL2
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2'-deoxyadenosine-3'-fluoro-3'-triphosphate
-
62% of the activity with ATP and deoxycytidine, 1.5% of the activity with ATP and deoxyadenosine
-
2'-deoxyadenosine-3'-fluoroara-3'-triphosphate
-
59% of the activity with ATP and deoxycytidine, 1% of the activity with ATP and deoxyadenosine
-
2'-deoxyadenosine-3'-triphosphate
-
73% of the activity with ATP and deoxycytidine, 6% of the activity with ATP and deoxyadenosine
-
3'-deoxyadenosine-2'-triphosphate
-
97% of the activity with ATP and deoxycytidine, 60% of the activity with ATP and deoxyadenosine
-
3'-deoxyadenosine-3'-fluoro-2'-triphosphate
-
66% of the activity with ATP and deoxycytidine, 4% of the activity with ATP and deoxyadenosine
-
3'-deoxyadenosine-3'-fluoroxylo-2'-triphosphate
-
62% of the activity with ATP and deoxycytidine, 4% of the activity with ATP and deoxyadenosine
-
additional information
-
triphosphate is active with 2'-deoxycytidine, 40% of the activity with ATP, not with 2'-deoxyadenosine, no activity with diphosphate as phosphate donor
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activation, can replace Mg2+
Mn2+
-
activation, can replace Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5'-amino-5'-deoxyadenosine
-
-
adenine arabinoside
-
-
adenosine
CHAPS
-
at critical micelle concentration
dADP
-
0.05 mM, 70% inhibition
dAMP
-
0.2 mM, 36% inhibition
dCDP
-
0.001 mM, 60% inhibition
dCMP
-
0.01 mM, 88% inhibition
deoxyadenosine
-
substrate ribavirin
deoxyguanosine
-
substrate deoxyadenosine
dGDP
-
0.2 mM, 46% inhibition
dGMP
-
0.5 mM, 32% inhibition
diphosphate
-
competitive inhibition
Hg2+
-
0.01 mM, 25% inhibition, 0.1 mM, almost complete inhibition
p-chloromercuribenzoate
-
0.0007 mM, 50% inhibition of ribavirin and deoxadenosine kinase activity
ribavirin
-
deoxyadenosine as substrate
Trypsin
-
substrates protect, end-products, i.e. dATP, dGTP or dCTP, protect corresponding active sites
-
additional information
-
not inhibited by adenosine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2'-O-Cyclocytidine
-
activation
5-Fluoro-2'-deoxycytidine
-
0.4 mM, 275% activation of deoxyadenosine phosphorylation, half-maximal activation at 0.028 mM
cytosine arabinoside
-
activation
deoxycytidine
-
30 mM, 584% activation of deoxyadenosine phosphorylation, half-maximal activation at 0.1 mM
deoxycytidine monophosphate
-
10 mM, 409% activation of deoxyadenosine phosphorylation, half-maximal activation at 1.6 mM
deoxyguanosine
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0335
2',2'-difluorodeoxycytidine
ratio kcat/Km is 5100 per s and M
0.192
2',2'-difluorodesoxycytidine
pH 8.0
0.0227 - 0.0332
2'-deoxyadenosine
0.027 - 0.0344
2'-deoxycytidine
0.146
2'-fluoro-arabinosyl adenine
-
0.007 - 0.06
ATP
0.021 - 0.022
CTP
0.0028 - 5
deoxyadenosine
0.027 - 0.18
deoxycytidine
0.019 - 0.021
deoxyguanosine
0.146
fludarabine
-
0.0335
gemcitabine
in 50 mM Tris/HCl, pH 8.0, 10 mM dithiothreitol, 2.5 mM ATP, 2.5 mM MgCl2, 3 mg/ml of bovine serum albumin and 0.5 mM CHAPS
0.11 - 0.44
MgATP2-
3.2
ribavirin
-
pH 7.5, 37C
0.074
Triphosphate
-
pH 7.5, 37C, with deoxycytidine
0.063 - 0.077
TTP
0.135 - 0.18
UTP
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.19
2',2'-difluorodeoxycytidine
Staphylococcus aureus
Q0H0H0
pH 8.0
1.82 - 3.7
2'-deoxyadenosine
0.56
2'-deoxycytidine
Staphylococcus aureus
Q0H0H0
pH 8.0
3.3
2'-fluoro-arabinosyl adenine
Dictyostelium discoideum
Q54YL2
-
0.84
ATP
Mycoplasma mycoides
-
pH 7.6, 37C, recombinant enzyme
0.62
CTP
Mycoplasma mycoides
-
pH 7.6, 37C, recombinant enzyme
1.22 - 3.7
deoxyadenosine
0.55 - 1.23
deoxycytidine
0.61
deoxyguanosine
Mycoplasma mycoides
-
pH 7.6, 37C, recombinant enzyme
3.3
fludarabine
Dictyostelium discoideum
Q54YL2
-
0.17
gemcitabine
Bacillus cereus
Q0H0H5
in 50 mM Tris/HCl, pH 8.0, 10 mM dithiothreitol, 2.5 mM ATP, 2.5 mM MgCl2, 3 mg/ml of bovine serum albumin and 0.5 mM CHAPS
0.0081 - 8.1
thymidine
1.64
TTP
Mycoplasma mycoides
-
pH 7.6, 37C, recombinant enzyme
0.36
UTP
Mycoplasma mycoides
-
pH 7.6, 37C, recombinant enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.035 - 0.057
adenosine
0.001 - 0.0014
dATP
0.97
deoxyadenosine
-
pH 7.5, 37C, substrate ribavirin
0.08 - 0.31
diphosphate
2.5
ribavirin
-
pH 7.5, 37C, substrate deoxyadenosine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0000045
-
in 30 mM HEPES-NaOH buffer (pH 7.6), 10 mM MgCl2, 1 mM DTT, 3.75 mM ATP and 0.055 mM deoxyadenosine
0.007
-
purified recombinant enzyme, substrates deoxycytidine and ATP
0.027
-
substrate 2'-deoxycytidine and ATP
0.06
-
substrate ribavarin
0.073
-
-
0.43
-
purified recombinant enzyme, substrates deoxyadenosine and ATP
0.54
-
substrate 2'-deoxyadenosine and ATP
2.3
-
dAdo/dCyd kinase in crude recombinant Escherichia coli cell extract, substrate 2'-deoxyadenosine or 2'-deoxycytidine
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.7 - 8.8
-
approx. half-maximal activity at pH 5.7, approx. 80% of maximal activity at pH 8.8
6 - 9
-
approx. 30% of maximal activity at pH 6.0 and pH 9.0, respectively
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27 - 50
-
approx. half-maximal activity at 27C and 50C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
low activity
Manually annotated by BRENDA team
-
very low activity
Manually annotated by BRENDA team
-
peripheral from adult, very low activity
Manually annotated by BRENDA team
-
low activity
Manually annotated by BRENDA team
-
low activity
Manually annotated by BRENDA team
-
low activity
Manually annotated by BRENDA team
-
peripheral from adult
Manually annotated by BRENDA team
-
low activity
Manually annotated by BRENDA team
-
suspension culture
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26200
SDS-PAGE
63000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
three-dimensional structural model based on crystal structure of Mycoplasma mycoides deoxyadenosine kinase
in complex with deoxyadenosine 5'-triphosphate and 2'-deoxycytidine 5'-triphosphate, as well as with products 2'-deoxycytidine 5'-phosphate and 2'-deoxycytidine 5'-diphosphate. Both deoxyadenosine 5'-triphosphate and 2'-deoxycytidine 5'-triphosphate bind to the enzyme in a feedback-inhibitory manner with the deoxyribonucleoside part in the deoxyribonucleoside binding site and the triphosphates in the P-loop. Superposition with human counterparts deoxyguanosine kinase and deoxycytidine kinase. Overall structures are very similar with a few amino acid alterations in the proximity of the active site; sitting drop vapour diffusion method using 24% (w/v) PEG 3350 and 0.2 M KSCN
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
half-life of deoxyadenosine kinase activity: 1.5 days
37
-
t1/2: 2 h, 1 mM ATP, 0.5 mM dTTP, 1 mM deoxyadenosine or 0.25 3-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate stabilize
42
-
first-order rate constant of inactivation: in the absence of dithiothreitol, 14.2/min, in the presence of 10 mM dithiothreitol, 6.82, substrate ribavirin
48
-
inactivation of deoxyadenosine kinase activity
50
-
80% loss of activity
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
dATP stabilizes
-
dithiothreitol protects against heat inactivation, glycerol and dithiothreitol increase stability during storage
-
glycerol, 50%, or dithiothreitol stabilizes
-
stability of deoxyadenosine kinase activity differs appreciably from deoxycytidine kinase activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 60% glycerol, at least 3 months, slight loss of activity
-
4C, t1/2: 84 h with and t1/2: 32 h without addition of bovine serum albumin
-
ATP, dTTP, deoxyadenosine or CHAPS stabilizes at 4C
-
dATP stabilizes during storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography
-
affinity chromatography on Cibacron Blue F3G-A
-
ammonium sulfate, Sephadex G-100, DEAE-cellulose, Sephadex G-75
-
glutathione Sepharose FF column chromatography; recombinant protein
recombinant dGK/dAK, dATP-Sepharose affinity chromatography
recombinant His-tagged enzyme, metal affinity chromatography
-
streptomycin sulfate, protamine sulfate, ammonium sulfate, Sephadex G-150
-
Superdex 75 column gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli; expression in Escherichia coli
expression in Escherichia coli
expression of dGK/dAK in Escherichia coli
gene yaaF of the yaaG/yaaF operon, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D78A
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mutation on both subunits of dGK/dAK, 0.2% of wild-type dAK activity
D78E
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mutation on both subunits of dGK/dAK, 0.2% of wild-type dAK activity
D78N
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mutation on both subunits of dGK/dAK, 0.2% of wild-type dAK activity
D84A
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mutation in dGK of dAK/dGK, increase of dAK activity
D84E
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mutation in dGK of dAK/dGK, increase of dAK activity
D84N
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mutation in dGK of dAK/dGK, increase of dAK activity
R79k
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2 types of mutants, one bears the mutation on both the dAK and dGK subunits while the other, called R79K:dGK, has the mutation on the dGK subunit only, strong increase in dAK activity, activation of dAK by deoxyguanosine is nearly eliminated in the case of R79K:dGK, while for the tandem mutated R79K mutant a 30% inhibition is observed, inhibition by dATP is reduced
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
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thymocytes lacking adenosine deaminase activity accumulate intracellular dATP and undergo apoptosis. Inhibition of both deoxycytidine kinase and adenosine kinase completely abrogates accumulation of dATP and significantly reduces the induction of apoptosis
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