Crystallization (Comment) | Organism |
---|---|
bifunctional D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase in complex with AMP and fructose 1,6-bisphosphate, to 2.3 A resolution. The allosteric inhibitor AMP and the substrate fructose 1,6-bisphosphate exhibit an unusual binding mode when in complex with D-fructose 1,6-bisphosphatase/sedoheptulose 1,7-bisphosphatase. AMP binds to the allosteric sites near the interface across the up/down subunit pairs C1C4 and C2C3 in the center of the tetramer, while fructose 1,6-bisphophate binds opposite to the interface between the horizontal subunit pairs C1C2 or C3C4. Formation of a disulfide linkage may occur between Cys75 and Cys99. The enzyme may be regulated through ligand binding and alteration of the structure of the enzyme complex | Synechocystis sp. |
Protein Variants | Comment | Organism |
---|---|---|
D198H | more than 10fold reduction in kcat value, reduction in IC50 value of AMP | Synechocystis sp. |
D200A | simultaneous decrease in substrate affinity and kcat value | Synechocystis sp. |
D97A | almost complete loss of activity | Synechocystis sp. |
E100A | almost complete loss of activity | Synechocystis sp. |
E225A | almost complete loss of activity | Synechocystis sp. |
E57A | almost complete loss of activity | Synechocystis sp. |
F309A | greatly reduced activity | Synechocystis sp. |
H215A | greatly reduced activity | Synechocystis sp. |
K134A | greatly reduced activity, reduction in IC50 value of AMP | Synechocystis sp. |
K29A | 2fold increase in activity | Synechocystis sp. |
N213A | greatly reduced activity | Synechocystis sp. |
R164A | greatly reduced activity | Synechocystis sp. |
R176A | simultaneous decrease in substrate affinity and kcat value | Synechocystis sp. |
R178G | greatly reduced activity | Synechocystis sp. |
R307A | greatly reduced activity | Synechocystis sp. |
R314A | greatly reduced activity | Synechocystis sp. |
T102A | more than 10fold reduction in kcat value, reduction in IC50 value of AMP | Synechocystis sp. |
Y131A | greatly reduced activity | Synechocystis sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
AMP | - |
Synechocystis sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.08 | - |
D-fructose 1,6-bisphosphate | mutant N213A, Hill coefficient 1.8, pH 8.0, 28°C | Synechocystis sp. | |
0.08 | - |
D-fructose 1,6-bisphosphate | mutant R314A, Hill coefficient 1.8, pH 8.0, 28°C | Synechocystis sp. | |
0.08 | - |
D-fructose 1,6-bisphosphate | wild-type, Hill coefficient 1.9, pH 8.0, 28°C | Synechocystis sp. | |
0.1 | - |
D-fructose 1,6-bisphosphate | mutant H215A, Hill coefficient 1.4, pH 8.0, 28°C | Synechocystis sp. | |
0.1 | - |
D-fructose 1,6-bisphosphate | mutant T102A, Hill coefficient 2.5, pH 8.0, 28°C | Synechocystis sp. | |
0.11 | - |
D-fructose 1,6-bisphosphate | mutant R164A, Hill coefficient 2.1, pH 8.0, 28°C | Synechocystis sp. | |
0.15 | - |
D-fructose 1,6-bisphosphate | mutant K29A, Hill coefficient 2.0, pH 8.0, 28°C | Synechocystis sp. | |
0.17 | - |
D-fructose 1,6-bisphosphate | mutant D198H, Hill coefficient 1.9, pH 8.0, 28°C | Synechocystis sp. | |
0.21 | - |
D-fructose 1,6-bisphosphate | mutant F309A, Hill coefficient 1.8, pH 8.0, 28°C | Synechocystis sp. | |
0.24 | - |
D-fructose 1,6-bisphosphate | mutant R307A, Hill coefficient 1.6, pH 8.0, 28°C | Synechocystis sp. | |
0.26 | - |
D-fructose 1,6-bisphosphate | mutant Y131A, Hill coefficient 1.6, pH 8.0, 28°C | Synechocystis sp. | |
0.5 | - |
D-fructose 1,6-bisphosphate | mutant K134A, Hill coefficient 2.2, pH 8.0, 28°C | Synechocystis sp. | |
0.61 | - |
D-fructose 1,6-bisphosphate | mutant R178A, Hill coefficient 1.1, pH 8.0, 28°C | Synechocystis sp. | |
1.32 | - |
D-fructose 1,6-bisphosphate | mutant D200A, Hill coefficient 1.4, pH 8.0, 28°C | Synechocystis sp. | |
3.2 | - |
D-fructose 1,6-bisphosphate | mutant R176A, Hill coefficient 1.2, pH 8.0, 28°C | Synechocystis sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Synechocystis sp. | P73922 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 1,6-bisphosphate + H2O | - |
Synechocystis sp. | D-fructose 6-phosphate + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
slr2094 | - |
Synechocystis sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.63 | - |
D-fructose 1,6-bisphosphate | mutant D200A, Hill coefficient 1.4, pH 8.0, 28°C | Synechocystis sp. | |
0.7 | - |
D-fructose 1,6-bisphosphate | mutant R176A, Hill coefficient 1.2, pH 8.0, 28°C | Synechocystis sp. | |
0.7 | - |
D-fructose 1,6-bisphosphate | mutant T102A, Hill coefficient 2.5, pH 8.0, 28°C | Synechocystis sp. | |
0.84 | - |
D-fructose 1,6-bisphosphate | mutant D198H, Hill coefficient 1.9, pH 8.0, 28°C | Synechocystis sp. | |
1.2 | - |
D-fructose 1,6-bisphosphate | mutant H215A, Hill coefficient 1.4, pH 8.0, 28°C | Synechocystis sp. | |
1.2 | - |
D-fructose 1,6-bisphosphate | mutant Y131A, Hill coefficient 1.6, pH 8.0, 28°C | Synechocystis sp. | |
1.8 | - |
D-fructose 1,6-bisphosphate | mutant K134A, Hill coefficient 2.2, pH 8.0, 28°C | Synechocystis sp. | |
2.3 | - |
D-fructose 1,6-bisphosphate | mutant R164A, Hill coefficient 2.1, pH 8.0, 28°C | Synechocystis sp. | |
2.6 | - |
D-fructose 1,6-bisphosphate | mutant R314A, Hill coefficient 1.8, pH 8.0, 28°C | Synechocystis sp. | |
5.2 | - |
D-fructose 1,6-bisphosphate | mutant F309A, Hill coefficient 1.8, pH 8.0, 28°C | Synechocystis sp. | |
5.2 | - |
D-fructose 1,6-bisphosphate | mutant N213A, Hill coefficient 1.8, pH 8.0, 28°C | Synechocystis sp. | |
5.2 | - |
D-fructose 1,6-bisphosphate | mutant R307A, Hill coefficient 1.6, pH 8.0, 28°C | Synechocystis sp. | |
7.1 | - |
D-fructose 1,6-bisphosphate | mutant R178A, Hill coefficient 1.1, pH 8.0, 28°C | Synechocystis sp. | |
10.5 | - |
D-fructose 1,6-bisphosphate | wild-type, Hill coefficient 1.9, pH 8.0, 28°C | Synechocystis sp. | |
21.3 | - |
D-fructose 1,6-bisphosphate | mutant K29A, Hill coefficient 2.0, pH 8.0, 28°C | Synechocystis sp. |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.015 | - |
mutant Y131A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
0.0214 | - |
mutant R176A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
0.027 | - |
mutant R178A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
0.028 | - |
mutant K29A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
0.034 | - |
wild-type, pH 8.0, 28°C | Synechocystis sp. | AMP | |
0.21 | - |
mutant N213A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
0.225 | - |
mutant D200A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
1.23 | - |
mutant D198H, pH 8.0, 28°C | Synechocystis sp. | AMP | |
1.241 | - |
mutant T102A, pH 8.0, 28°C | Synechocystis sp. | AMP | |
2.099 | - |
mutant K134A, pH 8.0, 28°C | Synechocystis sp. | AMP |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.21 | - |
D-fructose 1,6-bisphosphate | mutant R176A, pH 8.0, 28°C | Synechocystis sp. | |
0.48 | - |
D-fructose 1,6-bisphosphate | mutant D200A, pH 8.0, 28°C | Synechocystis sp. | |
3.6 | - |
D-fructose 1,6-bisphosphate | mutant K134A, pH 8.0, 28°C | Synechocystis sp. | |
4.7 | - |
D-fructose 1,6-bisphosphate | mutant Y131A, pH 8.0, 28°C | Synechocystis sp. | |
4.8 | - |
D-fructose 1,6-bisphosphate | mutant D198H, pH 8.0, 28°C | Synechocystis sp. | |
6.8 | - |
D-fructose 1,6-bisphosphate | mutant T102A, pH 8.0, 28°C | Synechocystis sp. | |
12 | - |
D-fructose 1,6-bisphosphate | mutant H215A, pH 8.0, 28°C | Synechocystis sp. | |
12 | - |
D-fructose 1,6-bisphosphate | mutant R178A, pH 8.0, 28°C | Synechocystis sp. | |
22 | - |
D-fructose 1,6-bisphosphate | mutant R164A, pH 8.0, 28°C | Synechocystis sp. | |
22 | - |
D-fructose 1,6-bisphosphate | mutant R307A, pH 8.0, 28°C | Synechocystis sp. | |
25 | - |
D-fructose 1,6-bisphosphate | mutant F309A, pH 8.0, 28°C | Synechocystis sp. | |
32 | - |
D-fructose 1,6-bisphosphate | mutant R314A, pH 8.0, 28°C | Synechocystis sp. | |
61 | - |
D-fructose 1,6-bisphosphate | mutant N213A, pH 8.0, 28°C | Synechocystis sp. | |
130 | - |
D-fructose 1,6-bisphosphate | wild-type, pH 8.0, 28°C | Synechocystis sp. | |
140 | - |
D-fructose 1,6-bisphosphate | mutant K29A, pH 8.0, 28°C | Synechocystis sp. |