Information on EC 3.6.1.13 - ADP-ribose diphosphatase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.6.1.13
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RECOMMENDED NAME
GeneOntology No.
ADP-ribose diphosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ADP-D-ribose + H2O = AMP + D-ribose 5-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphorous acid anhydride hydrolysis
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
purine metabolism
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Purine metabolism
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SYSTEMATIC NAME
IUBMB Comments
ADP-D-ribose ribophosphohydrolase
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CAS REGISTRY NUMBER
COMMENTARY hide
9024-83-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Uniprot
Manually annotated by BRENDA team
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UniProt
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
PCC 7002
SwissProt
Manually annotated by BRENDA team
enzyme displays both nicotinamide mononucleotide deamidase and ADP-ribose diphosphatase activities
UniProt
Manually annotated by BRENDA team
Thermus thermophilus HB8 / ATCC 27634 / DSM 579
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
NUDX7 contributes to 23% of the total diphosphohydrolase activity toward ADP-ribose under normal conditions, while under oxidative stress, the contribution of NUDX7 to the activity increases to 34%. Additionally, NUDX7 accounts for 53% of the total pyrophosphohydrolase activity toward NADH under normal conditions and the activity is increased by oxidative stress with NUDX7 contributing 57%
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,Nepsilon-etheno-ADP-ribose + H2O
1,Nepsilon-etheno-AMP-ribose + phosphate
show the reaction diagram
-
fluorogenic substrate
the product is converted to fluorescent 1,Nepsilon-etheno-adenosine by alkaline phosphatase for detection
-
?
2''-O-acetyl-ADP-ribose + H2O
AMP + 2-O-acetyl-D-ribose 5-phosphate
show the reaction diagram
-
-
-
-
?
2',3'-cAMP + H2O
3'AMP
show the reaction diagram
8-oxo-dADP + H2O
?
show the reaction diagram
-
-
-
-
?
8-oxo-dGDP + H2O
8-oxo-dGMP + phosphate
show the reaction diagram
-
-
-
-
?
8-oxo-dGDP + H2O
?
show the reaction diagram
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-
-
-
?
8-oxo-dGTP + H2O
?
show the reaction diagram
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the enzyme hardly acts on 8-oxo-dGTP
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-
?
ADP + H2O
AMP + phosphate
show the reaction diagram
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-
-
?
ADP-D-ribose + H2O
AMP + D-ribose 5-phosphate
show the reaction diagram
ADP-glucose + H2O
?
show the reaction diagram
ADP-glucose + H2O
AMP + alpha-D-glucose 1-phosphate
show the reaction diagram
ADP-mannose + H2O
AMP + D-mannose 1-phosphate
show the reaction diagram
ADP-ribose + H2O
AMP + D-ribose 5-phosphate
show the reaction diagram
ADP-ribose + H2O
AMP + ribose 5-phosphate
show the reaction diagram
ADPribose + H2O
AMP + D-ribose 5-phosphate
show the reaction diagram
ADPribose 2'-phosphate + H2O
adenosine 2',5'-diphosphate + D-ribose 5-phosphate
show the reaction diagram
-
-
-
-
?
cADP-ribose + H2O
N1-(5-phosphoribosyl)-AMP + phosphate
show the reaction diagram
CDP-choline + H2O
CMP + choline phosphate
show the reaction diagram
CDP-ethanolamine + H2O
CMP + ethanolamine phosphate
show the reaction diagram
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-
-
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CDP-glucose + H2O
CMP + glucose 5-phosphate
show the reaction diagram
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at 5% of the activity with ADPribose
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-
?
CDP-glycerol + H2O
CMP + glycerol phosphate
show the reaction diagram
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-
-
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CDP-ribose + H2O
CMP + D-ribose 5-phosphate
show the reaction diagram
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kcat/Km is 2.5% of the kcat/Km for ADP-ribose
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-
?
diadenosine 5',5''-diphosphate + H2O
?
show the reaction diagram
20% the activity with ADP-ribose
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-
?
FAD + H2O
?
show the reaction diagram
GDP-glucose + H2O
GMP + glucose
show the reaction diagram
7% of the activity with ADP-ribose
-
-
?
GDP-glucose + H2O
GMP + glucose 5-phosphate
show the reaction diagram
GDP-mannose + H2O
GMP + D-mannose 1-phosphate
show the reaction diagram
GDP-ribose + H2O
GMP + D-ribose 5-phosphate
show the reaction diagram
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kcat/Km is 3% of the kcat/Km for ADP-ribose
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-
?
IDP-ribose + H2O
IMP + D-ribose 5-phosphate
show the reaction diagram
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138% of the activity with ADPribose
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-
?
IDP-ribose + H2O
IMP + ribose 5-phosphate
show the reaction diagram
IDPribose + H2O
IMP + D-ribose 5-phosphate
show the reaction diagram
77% of the activity with ADPribose
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-
?
NAD+ + H2O
?
show the reaction diagram
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ADPRibase-Mn in presence of Mn2+, no activity with ADPRibase I and ADPRibase II
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-
?
NAD+ + H2O
AMP + NMN
show the reaction diagram
7% of the activity with ADP-ribose
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-
?
NADH + H2O
?
show the reaction diagram
NADH + H2O
AMP + NMNH
show the reaction diagram
NADPH + H2O
AMP + ?
show the reaction diagram
12% of the activity with ADP-ribose
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-
?
UDP-galactose + H2O
UDP + galactose
show the reaction diagram
7% of the activity with ADP-ribose
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-
?
UDP-glucose + H2O
UMP + glucose 5-phosphate
show the reaction diagram
UDP-mannose + H2O
UMP + D-mannose 1-phosphate
show the reaction diagram
additional information
?
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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
ADP-D-ribose + H2O
AMP + D-ribose 5-phosphate
show the reaction diagram
ADP-ribose + H2O
AMP + D-ribose 5-phosphate
show the reaction diagram
ADP-ribose + H2O
AMP + ribose 5-phosphate
show the reaction diagram
Q9UKK9
the function of the enzyme might be to remove free ADP-ribose arising from NAD+ and protein-bound poly- and non-enzymic protein glycation
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?
ADPribose + H2O
AMP + D-ribose 5-phosphate
show the reaction diagram
FAD + H2O
?
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,N6-ethenoNAD+
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2'-deoxyNAD+
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3'-deoxyNAD+
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3-acetylpyridineNAD+
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5-amino-3-(morpholin-4-yl)-1,2,3-oxadiazol-3-ium
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3-morpholinosynonimine increases Km and slightly decreases Vmax
8-BromoNAD+
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8-oxo-dGDP
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the ADP-ribose cleavage is competitively inhibited by 8-oxo-dGDP (68% relative cleavage efficiency at 0.01 mM)
8-oxo-dGMP
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the ADP-ribose cleavage is competitively inhibited by 8-oxo-dGMP (89% relative cleavage efficiency at 0.01 mM)
ADP
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potent noncompetitive inhibitor
ADP-ribose
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the 8-oxo-dGDP cleavage is competitively inhibited by ADP-ribose (22% relative cleavage efficiency at 0.005 mM)
fluoride
Mn2+
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above 0.05 mM
N-acetyl-p-benzoquinoneimine
nitroprusside
thionicotinamideNAD+
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additional information
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no inhibitory effect is observed with 8-oxo-dGMP
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
nitric oxide
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NO stimulates non-enzymatic ADP-ribosylation, at cysteine residues in the presence of reductant, of NUDT5 using ADP-ribose and consequently activates its ADPRase activity. ADPRase activity in J774 macrophage cells is increased by the treatment with SNP, an exogenous NO generator, or TNF-alpha/IFN-gamma, endogenous NO inducers. NO has a regulatory role, overview
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.045
2''-O-acetyl-ADP-ribose
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at pH 7.5 and 37°C
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0.76 - 7.6
2',3'-cAMP
0.0035 - 0.0038
8-oxo-dGDP
2.65 - 19
ADP
0.0209 - 0.3213
ADP-D-ribose
3.16
ADP-glucose
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25°C, pH 7.6, wild-type enzyme
0.083 - 0.154
ADP-mannose
0.0019 - 2.1
ADP-ribose
0.0004 - 0.37
ADPribose
0.46
ADPribose 2'-phosphate
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0.19 - 0.78
cADP-ribose
0.35 - 43
CDP-choline
3.9 - 31
CDP-ethanolamine
2 - 6.3
CDP-glycerol
1.36
CDP-ribose
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25°C, pH 7.6, wild-type enzyme
0.3 - 0.33
FAD
1.1
GDP-ribose
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25°C, pH 7.6, wild-type enzyme
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.78
2''-O-acetyl-ADP-ribose
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at pH 7.5 and 37°C
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0.11 - 83
2',3'-cAMP
0.2 - 1.8
ADP
0.0019 - 13.7
ADP-D-ribose
9.1
ADP-glucose
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25°C, pH 7.6, wild-type enzyme
0.00011 - 97
ADP-ribose
1.8 - 117
ADPribose
1.14
ADPribose 2'-phosphate
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-
0.0005 - 16
cADP-ribose
0.025 - 79
CDP-choline
2.1 - 125.2
CDP-ethanolamine
0.8 - 95.6
CDP-glycerol
3.36
CDP-ribose
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25°C, pH 7.6, wild-type enzyme
2.5 - 5
FAD
3.57
GDP-ribose
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25°C, pH 7.6, wild-type enzyme
additional information
ADP-ribose
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25°C, pH 7.6, mutant enzyme I19A
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
17.3
2''-O-acetyl-ADP-ribose
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at pH 7.5 and 37°C
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0.1 - 32
2',3'-cAMP
0.0011 - 0.66
ADP
0.0027 - 630
ADP-D-ribose
0.03 - 1000
ADP-ribose
0.0001 - 44
cADP-ribose
0.002 - 180
CDP-choline
0.007 - 32
CDP-ethanolamine
0.12 - 48
CDP-glycerol
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015 - 0.15
fluoride
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02
F-
Homo sapiens
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IC50: 0.02 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.439
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ADPRibase I
8.59
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-
12.81
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ADPRibase II
18.2
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ADPRibase-Mn
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
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activity at pH 8.0 is about 90% of the activity at pH 6.5 and at pH 9.0, activity with ADP-ribose
7 - 9
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for cleavage of ADP-ribose
7 - 9
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ADPRibase I and ADPRibase II
7.4 - 9
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with 2.5-5 mM Mg2+ or 0.1-0.25 mM Mn2+
10
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for cleavage of 8-oxo-dGDP
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 10
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pH 4.5: about 55% of maximal activity, pH 10.0: about 50% of maximal activity, activity with ADP-mannose
6 - 10
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pH 6.0: about 25% of maximal activity, pH 10.0: about 60% of maximal activity, activity with ADP-ribose
7 - 9
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linearly decreasing activity profile from pH 7 to pH 9, with activity at pH 9 being 50% of that at pH 7
7.5 - 10
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pH 7.5: about 60% of maximal activity with ADP-ribose, about 50% of maximal activity with ADP-mannose, pH 10.0: about 70% of maximal activity with activity with ADP-ribose or ADP-mannose
8 - 10
pH 8.0: about 40% of maximal activity, pH 10.0: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 75
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rate of hydrolysis of ADPribose is 15fold higher than at 37°C, incubations at higher temperatures are impractical because of the spontaneous hydrolysis of the substrate
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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Raji and Daudi
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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activity in leaves is lower than in stem and root, AtNUDT10; activity in roots is lower than in leaves and stem, AtNUDT6; AtNUDT2; AtNUDT7
Manually annotated by BRENDA team
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dermis and epidermis of the outer and the inner prepuce
Manually annotated by BRENDA team
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activity in leaves is lower than in stem and root, AtNUDT10; activity in roots is lower than in leaves and stem, AtNUDT6; AtNUDT2; AtNUDT7
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bdellovibrio bacteriovorus (strain ATCC 15356 / DSM 50701 / NCIB 9529 / HD100)
Bdellovibrio bacteriovorus (strain ATCC 15356 / DSM 50701 / NCIB 9529 / HD100)
Bdellovibrio bacteriovorus (strain ATCC 15356 / DSM 50701 / NCIB 9529 / HD100)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Mycobacterium tuberculosis (strain CDC 1551 / Oshkosh)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21800
2 * 21800, calculated and gel filtration
24300
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2 * 24300, native protein, mass spectrometry
26735
6 * 26735, calculation from nucleotide sequence
28000
6 * 28000, SDS-PAGE
31000
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gel filtration
31600
deduced from amino acid sequence
34000
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2 * 34000, SDS-PAGE with and without mercaptoethanol
35000
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gel filtration
36000 - 37000
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gel filtration
38000
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gel filtration
39121
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1 * 39121, calculation from nucleotide sequence
68000
-
gel filtration
85000
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dimeric thioredoxin fusion protein expressed in Escherichia coli, gel filtration
166000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
6 * 26735, calculation from nucleotide sequence; 6 * 28000, SDS-PAGE
homodimer
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monomer
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1 * 39121, calculation from nucleotide sequence
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
substrate-docking to zebrafish wild-type protein, and H97A mutant point to a role of His-97 in catalysis by orientation, and to a bidentate water bridging the dinuclear metal center as the potential nucleophile
gadolinium derivative, to 2.0 A resolution. The crystal structure of DR2204 consists of the conserved alpha/beta/alpha sandwich fold typical of Nudix hydrolases, the Nudix box, residues 94-115, holding the alpha1 helix sits between two loops accessible to the solvent, while the other two helices, alpha2 and alpha3, lie on the other side of the central beta-sheet and participate in dimer-interface formation
hanging-drop vapor diffusion at 18°C. The structure of the apo enzyme, the active enzyme and the complex with ADP-ribose are determined to 1.9 A, 2.7 A and 2.3 A, respectively. The Nudix motif residues, folded as a loop-helix-loop tailored for diphosphate hydrolysis, compose the catalytic center
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purified recombinant enzyme in complex with the product AMP and Mn2+ ions in its Nudix active site, sitting drop vapor diffusion method, 20°C, 0.001 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 0.1 M Tris, pH 7.5, 200 mM MgCl2, and 19% PEG 3350, with or without 30 mM AMP, equilibration against the reservoir, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement method
complexed with 8-oxo-dGMP, 8-oxo-dGDP and 8-oxo-dADP, hanging drop vapor diffusion method, using 0.8 M NaH2PO4/1.2M K2HPO4 and 0.1 M acetate (pH 4.5), and 0.2M ammonium acetate, 35-40% (w/v) polyethylene glycol 3350 and 0.1 M sodium citrate (pH 6.2)
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in apo form, in complex with ADP-D-ribose, and in complex with AMP with bound Mg2+, hanging drop vapor diffusion method, using 160 mM sodium acetate (pH 5.5), and 25% (w/v) 2-methyl-2,4-pentanediol, or 300 mM di-ammonium hydrogen citrate, 6% (w/v) n-propanol and 15% (w/v) polyethylene glycol 3350, or 200 mM sodium acetate, 100 mM Tris-HCl (pH 8.0) and 30% (w/v) polyethylene glycol 4000
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in complex with alpha,beta-methyleneadenosine diphosphoribose and 3 Mg2+ ions, hanging drop vapor diffusion method, using 250 mM sodium acetate, 100 mM Tris-HCl, pH 8.0, and 29% (w/v) polyethylene glycol 4000; purified recombinant wild-type and truncated mutant NUDT5 in complex with a non-hydrolyzable ADPR analogue, alpha,beta-methyleneadenosine diphosphoribose, and three Mg2+ ions representing the transition state of the enzyme during catalysis, 20 mg/ml protein is incubated with 5 mM AMPCPR and 10 mM MgCl2 at 4 °C overnight, followed by hanging drop vapour diffusion method, wild-type enzyme in complex with AMPCPR, and truncation mutant DELTAhNUDT5 in complex with AMPCPR and Mg2+, 4 °C, mixing of equal volumes of the protein solution and the reservoir solution containing 250 mM NaAc, 100 mM Tris-HCl, pH 8.0, and 29% PEG 4000, ingle crystals of the plate-shape morphology grow after 1 month, X-ray diffraction structure determination and anaylsis at 2.0 A resolution, molecular modelling
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substrate docking on a homology model suggests possible interactions of ADP-ribose with seven residues located, with one exception (Cys253), either within the metallo-dependent phosphatases signature (Gln27, Asn110, His111), or in unique structural regions of the ADPRibase-Mn family: s2s3 (Phe37 and Arg43) and h7h8 (Phe210), around the active site entrance. Residue Phe37 is needed for ADP-ribose preference without catalytic effect. Arg43 is essential for catalysis. Cys253 is hindering for cADPR phosphohydrolase
purified recombinant His-tagged enzyme in complex with co-purified NAD and diphosphate complexed in the NadM-domain active site, and with ADPR substrate complexed in the Nudix-domain, hanging drop vapor diffusion method, 0.0015 ml of 15 mg/ml protein solution is mixed with an equal volume of reservoir solution containing 100 mM Tris, pH 7.5, and 1.5 M Li2SO4, 20°C, 3 days to 2 weeks, X-ray diffraction structure determination and analysis at 2.6 A resolution, selenomethionyl MAD phasing method
crystallized in absence or presence of ADP-ribose by hanging-drop vapour-diffusion method. 1.5 A resolution from the apo form using synchrotron radiation and 2.0 A resolution from the complexed form. Both crystals belong to space group P3(1)21 or P3(2)21 and contain one molecule in the asymmetric unit
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in complex with alpha,beta-methyleneadenosine diphosphoribose, sitting drop vapor diffusion method, using 18% (w/v) PEG 4000, 0.1 M sodium acetate buffer pH 5.3, 20% (w/v) glycerol, 0.2 M ammonium sulfate, at 20°C
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Ndx2 alone and in complex with Mg2+, with Mg2+ and AMP, and with Mg2+ and a nonhydrolyzable ADPR analogue, hanging-drop vapor diffusion method, 20 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 100 mM KCl, 0.001 ml of protein solution is mixed with the equal volume of reservoir solution and equilibrated against the reservoir, containing 0.1 M MES, pH 6.5, 0.16 M sodium acetate or magnesium acetate for the complexed enzyme, 14% PEG 8000, and 20% glycerol, at 20°C, soaking of crystals in 50 mM KAu(CN)2, X-ray diffraction structure determination and anaylsis at 2.0 A resolution, MAD phasing, model building, and refinement
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to 2.16 A resolution. The crystal structure shows an unusual asymmetric dimer, with three domains for each chain. The C-terminal domain harbors the nicotinamide mononucleotide deamidase activity. The N-terminal domain belongs to the COG1058 family and is associated with the ADP-ribose diphosphatase activity. The mechanism for the ADP-ribose diphosphatase reaction involves a rotation of the COG1058 domain dimer as part of the reaction cycle
Zn2+-bound enzyme, binary complex with ADPribose, ternary complex with Zn2+ and ADPribose, ternary complex with Gd3+ and ADPribose, product complex with AMP and Mg2+, product complex with ribose 5'-phosphate and Zn2+, mutant enzyme E82Q with ligands Mg2+ and SO42-, mutant enzyme E86Q with ligands Mg2+ and ADPribose, mutant enzyme E82Q with ligands Zn2+ and SO42-, mutant enzyme E86Q with ligands Zn2+ and ADPribose
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
-
5 min, retains full activity
85
-
5 min, 30% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, no loss of activity after months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
acetaminophen-treated animals and controls; ADPribose pyrophosphatase I; partial
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ADP-ribose pyrophosphatase Sll1054; ADP-ribose pyrophosphatase Slr0920; ADP-ribose pyrophosphatase Slr1134
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ADPRibaseI, ADPribase II and ADPRibase-Mn
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ammonium sulfate precipitation, Toyopearl Phenyl-650M column chromatography, and Q Sepharose column chromatography
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AtNUDT10; AtNUDT2; AtNUDT6; AtNUDT7
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Ni-NTA column chromatography and Q-Sepharose column chromatography
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Ni-Sepharose 6 column chromatography, HisTrap column chromatography, and Superdex 75 gel filtration
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partial
recombinant enzyme
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, the tag is cleaved off by TEV protease, and the detagged protein is further purified by anion exchange chromatography
recombinant His-tagged enzyme from Escherichia coli strain Bl21 by nickel affinity and anion exchange chromatography, and gel filtration
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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recombinant His6-tagged NUDT5 from Escherichia coli strain BL21 by nickel affinity chromatography
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recombinant Ndx2 from Escherichia coli strain BL21(DE3) by heat treatment at 70°C for 15 min and ion exchange chromatography
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Talon resin column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; overexpression of wild-type and mutant enzymes
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expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli M15 cells
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expression as a thioredoxin fusion protein in Escherichia coli
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expression in Escherichia coli
expression in Escherichia coli, ADP-ribose pyrophosphatase Sll1054; expression in Escherichia coli, ADP-ribose pyrophosphatase Slr0920; expression in Escherichia coli, ADP-ribose pyrophosphatase Slr1134
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expression in Escherichia coli, AtNUDT10; expression in Escherichia coli, AtNUDT2; expression in Escherichia coli, AtNUDT6; expression in Escherichia coli, AtNUDT7
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expression of GST-tagged nezyme in Escherichia coli
expression of Ndx2 in Escherichia coli strain BL21(DE3)
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expression of ORF38 as His6-tagged protein in Escherichia coli strain Bl21(DE3)
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expression of the His-tagged enzyme in Escherichia coli strain Bl21
gene NUDT5, expression in murine J774A.1 macrophage cells, expression of His6-tagged NUDT5 in Escherichia coli strain BL21
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hexahistidine-tagged recombinant enzyme expressed in Escherichia coli
overproduction in Escherichia coli
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
oxidative stress caused by 0.003 mM paraquat for 7 days causes an increase in the total ADP-ribose diphosphohydrolase activity of NUDX7
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H97A
mutant shows 60fold decrease in activity for substrates ADP-ribose and ADP and 300–500fold for CDP-alcohols. For H97A, 2',3'-cAMP is a better substrate than ADP-ribose
C139A
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site-directed mutagenesis, mutation causes a 2.1fold increase in Km for ADP-ribose but has no effect on kcat compared to the wild-type enzyme; the mutant shows reduced catalytic efficiency compared to the wild type enzyme
C253A
mutant displays a tenfold increased efficiency for cADP-ribose, with no or modest effect on the other substrates
D133A
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site-directed mutagenesis, mutation causes a 4.0fold increase in Km for ADP-ribose but has no effect on kcat compared to the wild-type enzyme; the mutant shows reduced catalytic efficiency compared to the wild type enzyme
D133N
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site-directed mutagenesis, mutation causes a 2.1fold increase in Km for ADP-ribose but has no effect on kcat compared to the wild-type enzyme; the mutant shows reduced catalytic efficiency compared to the wild type enzyme
D164A
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the mutant shows reduced catalytic efficiency compared to the wild type enzyme
D164N
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the mutant shows increased catalytic efficiency compared to the wild type enzyme
E112Q
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the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme
E115Q
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the mutant shows reduced catalytic efficiency compared to the wild type enzyme
E116Q
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the mutant shows reduced catalytic efficiency compared to the wild type enzyme
E166Q
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the mutant shows reduced catalytic efficiency compared to the wild type enzyme
E93Q
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site-directed mutagenesis, mutation causes a 1.8fold increase in Km for ADP-ribose but has no effect on kcat compared to the wild-type enzyme; the mutant shows reduced catalytic efficiency compared to the wild type enzyme
F210A
lowers 40-70fold the catalytic efficiency for ADP-ribose, CDP-choline and 2',3'-cAMP hydrolysis, and 500fold for cADP-ribose
F37A
19fold increased Km for ADP-ribose, with only a 2-3fold increase of the CDP-choline and 2',3'-cAMP Km values
F37A/L196A
mutation lessens the relative preference for ADP-ribose versus cADP-ribose
F37A/L196F
mutation lessens the relative preference for ADP-ribose versus cADP-ribose
F37A/L196F/C253A
cyclic ADP-ribose is the best substrate for the mutant
F37Y
similar kinetic parameters as the wild type
H111A
marked efficiency decrease with all substrates except 2',3'-cAMP
H111N
marked efficiency decrease with all substrates except 2',3'-cAMP
L196A