Protein Variants | Comment | Organism |
---|---|---|
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 |
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 |
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 |
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 |
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 |
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 |
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 |
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 | Comment | Organism | Structure |
---|---|---|---|
F- | - |
Lupinus angustifolius |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | |
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 | Lupinus angustifolius | |
0.5 | - |
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 | |
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 | |
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 | |
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 | |
2.5 | - |
Ap4A | pH 8.0, 25°C, wild-type enzyme | Lupinus angustifolius | |
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 | |
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] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
19120 | - |
wild-type enzyme, MALDI-MS | Lupinus angustifolius |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Lupinus angustifolius | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(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 | Lupinus angustifolius | ? | - |
? | |
Ap4A + H2O | - |
Lupinus angustifolius | ? | - |
? | |
diadenosine 5',5''-P1,P4-tetraphosphate + H2O | - |
Lupinus angustifolius | ATP + AMP | - |
? |
Synonyms | Comment | Organism |
---|---|---|
(asymmetrical) diadenosine 5',5''-P1,P4-tetraphosphate hydrolase | - |
Lupinus angustifolius |
Ap4A hydrolase | - |
Lupinus angustifolius |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | |
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 | |
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 | |
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 | |
3.7 | - |
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 | |
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 | |
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 | |
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 | |
40.8 | - |
Ap4A | pH 8.0, 25°C, wild-type enzyme | Lupinus angustifolius |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
F- | pH 8.0, 25°C, wild-type enzyme | Lupinus angustifolius | |
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 | |
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 | |
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 | |
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 | |
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 | |
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 |