BRENDA - Enzyme Database

Characterization of active-site residues in diadenosine tetraphosphate hydrolase from Lupinus angustifolius

Maksel, D.; Gooley, P.R.; Swarbrick, J.D.; Guranowski, A.; Gange, C.; Blackburn, G.M.; Gayler, K.R.; Biochem. J. 357, 399-405 (2001)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
3.6.1.17
E122Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 20% of the wild-type value, turnover-number is 50% of the wild-type value. 2.3fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E125Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser. KM-value is 0.7% of the wild-type value, turnover-number is 56% of the wild-type value. Cleavage of (2'-pdA)AppppA to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP as with wild-type enzyme, the ratio of ATP released is increased compared to wild-type. 83fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E55Q
fusion protein containing and N-terminal extension of Gly-Pro-Leu-Gly-Ser. KM-value is 4.8% of the wild-type value, turnover-number is 0.01% of the wild-type value. Activity with (2'-pdA)AppppA is to low to be detected. More than 330fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E58D
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 38.4% of the wild-type value, turnover-number is 9% of the wild-type value. Cleavage of (2'-pdA)AppppA to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP as with wild-type enzyme, the ratio of ATP released is increased compared to wild-type. 16.7fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E58Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 80% of the wild-type value, turnover-number is 43.9% of the wild-type value. 6.7fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E59D
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 4fold higher than the wild-type value, turnover-number is 0.02% of the wild-type value. Activity with (2'-pdA)AppppA is to low to be detected. 2fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E59Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 2.4fold higher than the wild-type value, turnover-number is 0.00076% of the wild-type value. Activity with (2'-pdA)AppppA is to low to be detected
Lupinus angustifolius
3.6.1.17
R54Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. Cleavage of (2'-pdA)AppppA to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP as with wild-type enzyme, the ratio of ATP released is increased compared to wild-type. 330fold reduction in sensitivity to fluoride
Lupinus angustifolius
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.6.1.17
F-
-
Lupinus angustifolius
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.6.1.17
0.12
-
Ap4A
pH 8.0, 25°C, mutant enzyme E55Q fusion protein containing and N-terminal extension of Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
0.5
-
Ap4A
pH 8.0, 25°C, mutant enzyme E122Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys; pH 8.0, 25°C, mutant enzyme R54Q fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.96
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58D fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
1.4
-
Ap4A
pH 8.0, 25°C, mutant enzyme E125Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
2
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
2.5
-
Ap4A
pH 8.0, 25°C, wild-type enzyme
Lupinus angustifolius
3.6.1.17
5.9
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
10
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59D, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.6.1.17
19120
-
wild-type enzyme, MALDI-MS
Lupinus angustifolius
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.6.1.17
Lupinus angustifolius
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.6.1.17
(2'-pdA)AppppA + H2O
cleavage to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP occurs with a ratio of 60:40 with the wild-type enzyme and the mutant enzymes R54Q, E58D and E125Q. With the mutants R54Q, E58D and E125Q, releasing ATP is increased to 70% of total hydrolysis. Activity of mutants E55Q, E59D and E59Q is to low to be detected
654489
Lupinus angustifolius
?
-
-
-
?
3.6.1.17
diadenosine 5',5''-P1,P4-tetraphosphate + H2O
-
654489
Lupinus angustifolius
ATP + AMP
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.6.1.17
0.00031
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.005
-
Ap4A
pH 8.0, 25°C, mutant enzyme E55Q fusion protein containing and N-terminal extension of Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
0.011
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59D, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.29
-
Ap4A
pH 8.0, 25°C, mutant enzyme E125Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
3.7
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58D fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
4.2
-
Ap4A
pH 8.0, 25°C, mutant enzyme R54Q fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
17.9
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
20.4
-
Ap4A
pH 8.0, 25°C, mutant enzyme E122Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
40.8
-
Ap4A
pH 8.0, 25°C, wild-type enzyme
Lupinus angustifolius
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.6.1.17
0.003
-
F-
pH 8.0, 25°C, wild-type enzyme
Lupinus angustifolius
3.6.1.17
0.006
-
F-
pH 8.0, 25°C, mutant enzyme E59D, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.007
-
F-
pH 8.0, 25°C, mutant enzyme E122Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.02
-
F-
pH 8.0, 25°C, mutant enzyme E58Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.05
-
F-
pH 8.0, 25°C, mutant enzyme E58D fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.25
-
F-
pH 8.0, 25°C, mutant enzyme E125Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
1
-
F-
pH 8.0, 25°C, mutant enzyme R54Q fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
3.6.1.17
E122Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 20% of the wild-type value, turnover-number is 50% of the wild-type value. 2.3fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E125Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser. KM-value is 0.7% of the wild-type value, turnover-number is 56% of the wild-type value. Cleavage of (2'-pdA)AppppA to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP as with wild-type enzyme, the ratio of ATP released is increased compared to wild-type. 83fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E55Q
fusion protein containing and N-terminal extension of Gly-Pro-Leu-Gly-Ser. KM-value is 4.8% of the wild-type value, turnover-number is 0.01% of the wild-type value. Activity with (2'-pdA)AppppA is to low to be detected. More than 330fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E58D
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 38.4% of the wild-type value, turnover-number is 9% of the wild-type value. Cleavage of (2'-pdA)AppppA to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP as with wild-type enzyme, the ratio of ATP released is increased compared to wild-type. 16.7fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E58Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 80% of the wild-type value, turnover-number is 43.9% of the wild-type value. 6.7fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E59D
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 4fold higher than the wild-type value, turnover-number is 0.02% of the wild-type value. Activity with (2'-pdA)AppppA is to low to be detected. 2fold reduction in sensitivity to fluoride
Lupinus angustifolius
3.6.1.17
E59Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. KM-value is 2.4fold higher than the wild-type value, turnover-number is 0.00076% of the wild-type value. Activity with (2'-pdA)AppppA is to low to be detected
Lupinus angustifolius
3.6.1.17
R54Q
fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys. Cleavage of (2'-pdA)AppppA to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP as with wild-type enzyme, the ratio of ATP released is increased compared to wild-type. 330fold reduction in sensitivity to fluoride
Lupinus angustifolius
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.6.1.17
F-
-
Lupinus angustifolius
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.6.1.17
0.003
-
F-
pH 8.0, 25°C, wild-type enzyme
Lupinus angustifolius
3.6.1.17
0.006
-
F-
pH 8.0, 25°C, mutant enzyme E59D, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.007
-
F-
pH 8.0, 25°C, mutant enzyme E122Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.02
-
F-
pH 8.0, 25°C, mutant enzyme E58Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.05
-
F-
pH 8.0, 25°C, mutant enzyme E58D fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.25
-
F-
pH 8.0, 25°C, mutant enzyme E125Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
1
-
F-
pH 8.0, 25°C, mutant enzyme R54Q fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.6.1.17
0.12
-
Ap4A
pH 8.0, 25°C, mutant enzyme E55Q fusion protein containing and N-terminal extension of Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
0.5
-
Ap4A
pH 8.0, 25°C, mutant enzyme E122Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys; pH 8.0, 25°C, mutant enzyme R54Q fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.96
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58D fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
1.4
-
Ap4A
pH 8.0, 25°C, mutant enzyme E125Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
2
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
2.5
-
Ap4A
pH 8.0, 25°C, wild-type enzyme
Lupinus angustifolius
3.6.1.17
5.9
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
10
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59D, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.6.1.17
19120
-
wild-type enzyme, MALDI-MS
Lupinus angustifolius
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
(2'-pdA)AppppA + H2O
cleavage to produce ATP and 2'-deoxyadenylated AMP rather than 2'-deoxyadenylated ATP + AMP occurs with a ratio of 60:40 with the wild-type enzyme and the mutant enzymes R54Q, E58D and E125Q. With the mutants R54Q, E58D and E125Q, releasing ATP is increased to 70% of total hydrolysis. Activity of mutants E55Q, E59D and E59Q is to low to be detected
654489
Lupinus angustifolius
?
-
-
-
?
3.6.1.17
diadenosine 5',5''-P1,P4-tetraphosphate + H2O
-
654489
Lupinus angustifolius
ATP + AMP
-
-
-
?
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
0.00031
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.005
-
Ap4A
pH 8.0, 25°C, mutant enzyme E55Q fusion protein containing and N-terminal extension of Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
0.011
-
Ap4A
pH 8.0, 25°C, mutant enzyme E59D, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
0.29
-
Ap4A
pH 8.0, 25°C, mutant enzyme E125Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser
Lupinus angustifolius
3.6.1.17
3.7
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58D fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
4.2
-
Ap4A
pH 8.0, 25°C, mutant enzyme R54Q fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
17.9
-
Ap4A
pH 8.0, 25°C, mutant enzyme E58Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
20.4
-
Ap4A
pH 8.0, 25°C, mutant enzyme E122Q, fusion protein containing and N-terminal extension Gly-Pro-Leu-Gly-Ser-Pro-Asn-Cys
Lupinus angustifolius
3.6.1.17
40.8
-
Ap4A
pH 8.0, 25°C, wild-type enzyme
Lupinus angustifolius