BRENDA - Enzyme Database

Characterisation of a bis(5-nucleosyl)-tetraphosphatase (asymmetrical) from Drosophila melanogaster

Winward, L.; Whitfield, W.G.; Woodman, T.J.; McLennan, A.G.; Safrany, S.T.; Int. J. Biochem. Cell Biol. 39, 943-954 (2007)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.6.1.17
DNA and amino acid sequence determination and analysis, sequence comparison, expression of His-tagged Apf in Spodoptera frugiperda Sf21 cells, expression of EGFP-tagged enzyme in Drosophila melanbogaster Mel-2 cells
Drosophila melanogaster
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.6.1.17
fluoride
fluoride potently inhibits diadenosine tetraphosphate hydrolysis in the presence of Mg2+, whereas it is ineffective in the presence of Zn2+, because inhibition involves a specific, MgF3--containing transition state analogue complex, overview
Drosophila melanogaster
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.6.1.17
additional information
-
additional information
kinetics strongly depend on pH and cation, overview
Drosophila melanogaster
3.6.1.17
0.009
-
5',5'-diadenosine tetraphosphate
pH 6.5, 1 mm Zn2+
Drosophila melanogaster
3.6.1.17
0.012
-
5',5'-diadenosine tetraphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
3.6.1.17
0.015
-
5',5'-diadenosine hexaphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.6.1.17
nucleus
association with euchromatin and facultative heterochromatin
Drosophila melanogaster
5634
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.6.1.17
Co2+
the enzyme is dependent on divalent cations
Drosophila melanogaster
3.6.1.17
Mg2+
the enzyme is dependent on divalent cations, preferred cation, best at 3-10 mM
Drosophila melanogaster
3.6.1.17
Mn2+
the enzyme is dependent on divalent cations
Drosophila melanogaster
3.6.1.17
additional information
no activation by Cu2+, Fe2+, Ni2+ or Ca2+
Drosophila melanogaster
3.6.1.17
Zn2+
the enzyme is dependent on divalent cations, high activity
Drosophila melanogaster
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.6.1.17
5',5'-diadenosine tetraphosphate + H2O
Drosophila melanogaster
-
ATP + AMP
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.6.1.17
Drosophila melanogaster
Q4V6M1
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.6.1.17
recombinant His-tagged Apf from Spodoptera frugiperda Sf21 cells by nickel affinity chromatography with removal of the His-tag
Drosophila melanogaster
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.6.1.17
additional information
patterns of Apf expression in Drosophila melanogaster tissues, semi-quantitative RT-PCR, highest expression levels in embryos and adult females, overview
Drosophila melanogaster
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.6.1.17
additional information
-
-
Drosophila melanogaster
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.6.1.17
5',5'-diadenosine hexaphosphate + H2O
diadenosine hexaphosphate is efficiently hydrolysed to ATP only at pH 7.5 with 20 mM Mg2+
687119
Drosophila melanogaster
2 ATP
-
-
-
?
3.6.1.17
5',5'-diadenosine pentaphosphate + H2O
-
687119
Drosophila melanogaster
ATP + ADP
-
-
-
?
3.6.1.17
5',5'-diadenosine tetraphosphate + H2O
-
687119
Drosophila melanogaster
ATP + AMP
-
-
-
?
3.6.1.17
5',5'-diadenosine tetraphosphate + H2O
best substrate
687119
Drosophila melanogaster
ATP + AMP
-
-
-
?
3.6.1.17
adenosine-5'-pentaphospho-5'-guanosine + H2O
-
687119
Drosophila melanogaster
?
-
-
-
?
3.6.1.17
adenosine-5'-tetraphospho-5'-guanosine + H2O
-
687119
Drosophila melanogaster
?
-
-
-
?
3.6.1.17
additional information
Apf always produces an NTP product, with substrate preference depending on pH and divalent ion
687119
Drosophila melanogaster
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
3.6.1.17
More
the enzyme contains a Nudix motif
Drosophila melanogaster
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.6.1.17
25
40
broad optimum
Drosophila melanogaster
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
3.6.1.17
20
60
65% of maximal activity at 52°C
Drosophila melanogaster
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.6.1.17
60
-
20 min, 55% remaining activity
Drosophila melanogaster
3.6.1.17
95
-
20 min, 1 mM Mn2+, a little amount of activity is remaining
Drosophila melanogaster
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.6.1.17
4
-
5',5'-diadenosine hexaphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
3.6.1.17
13
-
5',5'-diadenosine tetraphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
3.6.1.17
43
-
5',5'-diadenosine tetraphosphate
pH 6.5, 1 mM Zn2+
Drosophila melanogaster
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.6.1.17
6.5
7.5
activity depends on the substrate and the divalent cation
Drosophila melanogaster
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
3.6.1.17
0.02
-
in presence of Mg2+
Drosophila melanogaster
fluoride
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.6.1.17
DNA and amino acid sequence determination and analysis, sequence comparison, expression of His-tagged Apf in Spodoptera frugiperda Sf21 cells, expression of EGFP-tagged enzyme in Drosophila melanbogaster Mel-2 cells
Drosophila melanogaster
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
3.6.1.17
0.02
-
in presence of Mg2+
Drosophila melanogaster
fluoride
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.6.1.17
fluoride
fluoride potently inhibits diadenosine tetraphosphate hydrolysis in the presence of Mg2+, whereas it is ineffective in the presence of Zn2+, because inhibition involves a specific, MgF3--containing transition state analogue complex, overview
Drosophila melanogaster
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.6.1.17
additional information
-
additional information
kinetics strongly depend on pH and cation, overview
Drosophila melanogaster
3.6.1.17
0.009
-
5',5'-diadenosine tetraphosphate
pH 6.5, 1 mm Zn2+
Drosophila melanogaster
3.6.1.17
0.012
-
5',5'-diadenosine tetraphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
3.6.1.17
0.015
-
5',5'-diadenosine hexaphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.6.1.17
nucleus
association with euchromatin and facultative heterochromatin
Drosophila melanogaster
5634
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.6.1.17
Co2+
the enzyme is dependent on divalent cations
Drosophila melanogaster
3.6.1.17
Mg2+
the enzyme is dependent on divalent cations, preferred cation, best at 3-10 mM
Drosophila melanogaster
3.6.1.17
Mn2+
the enzyme is dependent on divalent cations
Drosophila melanogaster
3.6.1.17
additional information
no activation by Cu2+, Fe2+, Ni2+ or Ca2+
Drosophila melanogaster
3.6.1.17
Zn2+
the enzyme is dependent on divalent cations, high activity
Drosophila melanogaster
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.6.1.17
5',5'-diadenosine tetraphosphate + H2O
Drosophila melanogaster
-
ATP + AMP
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.6.1.17
recombinant His-tagged Apf from Spodoptera frugiperda Sf21 cells by nickel affinity chromatography with removal of the His-tag
Drosophila melanogaster
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.6.1.17
additional information
patterns of Apf expression in Drosophila melanogaster tissues, semi-quantitative RT-PCR, highest expression levels in embryos and adult females, overview
Drosophila melanogaster
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.6.1.17
additional information
-
-
Drosophila melanogaster
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.6.1.17
5',5'-diadenosine hexaphosphate + H2O
diadenosine hexaphosphate is efficiently hydrolysed to ATP only at pH 7.5 with 20 mM Mg2+
687119
Drosophila melanogaster
2 ATP
-
-
-
?
3.6.1.17
5',5'-diadenosine pentaphosphate + H2O
-
687119
Drosophila melanogaster
ATP + ADP
-
-
-
?
3.6.1.17
5',5'-diadenosine tetraphosphate + H2O
-
687119
Drosophila melanogaster
ATP + AMP
-
-
-
?
3.6.1.17
5',5'-diadenosine tetraphosphate + H2O
best substrate
687119
Drosophila melanogaster
ATP + AMP
-
-
-
?
3.6.1.17
adenosine-5'-pentaphospho-5'-guanosine + H2O
-
687119
Drosophila melanogaster
?
-
-
-
?
3.6.1.17
adenosine-5'-tetraphospho-5'-guanosine + H2O
-
687119
Drosophila melanogaster
?
-
-
-
?
3.6.1.17
additional information
Apf always produces an NTP product, with substrate preference depending on pH and divalent ion
687119
Drosophila melanogaster
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.6.1.17
More
the enzyme contains a Nudix motif
Drosophila melanogaster
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.6.1.17
25
40
broad optimum
Drosophila melanogaster
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
3.6.1.17
20
60
65% of maximal activity at 52°C
Drosophila melanogaster
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.6.1.17
60
-
20 min, 55% remaining activity
Drosophila melanogaster
3.6.1.17
95
-
20 min, 1 mM Mn2+, a little amount of activity is remaining
Drosophila melanogaster
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.6.1.17
4
-
5',5'-diadenosine hexaphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
3.6.1.17
13
-
5',5'-diadenosine tetraphosphate
pH 7.5, 20 mM Mg2+
Drosophila melanogaster
3.6.1.17
43
-
5',5'-diadenosine tetraphosphate
pH 6.5, 1 mM Zn2+
Drosophila melanogaster
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.6.1.17
6.5
7.5
activity depends on the substrate and the divalent cation
Drosophila melanogaster