Cloned (Comment) | Organism |
---|---|
gene pfkC, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Pyrococcus horikoshii |
gene pfkC, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Methanocaldococcus jannaschii |
phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | uncultured bacterium |
Crystallization (Comment) | Organism |
---|---|
purified enzyme mutant E72A, hanging drop vapor diffusion method, mixing of 0.002 ml of 8 mg/ml protein in 25 mM HEPES/NaOH, pH 7.8, 30 mM MgCl2, 20 mM fructose 6-phosphate, and 20 mM AMP with 0.002 ml of reservoir solution containing 18% PEG 3350 and 0.15 M NaI, 3 days at 18°C, X-ray diffraction structure determination and analysis at 2.61 A resolution, molecular replacement using the structure of Pyrococcus horikoshii ADP-PFK (PDB ID 3DRW) split into small and large domains as search models | uncultured bacterium |
Protein Variants | Comment | Organism |
---|---|---|
E72A | site-directed mutagenesis, the mutant shows reduced glucokinase and phosphofructokinase activities compared to wild-type enzyme | uncultured bacterium |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | enzyme PhPFK is not significantly inhibited by D-glucose | Pyrococcus horikoshii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0063 | - |
D-fructose 6-phosphate | pH 6.5, 40°C | Methanocaldococcus jannaschii | |
0.056 | - |
D-fructose 6-phosphate | pH 6.5, 40°C | Pyrococcus horikoshii | |
0.119 | - |
D-fructose 6-phosphate | pH 6.5, 40°C, wild-type enzyme | uncultured bacterium | |
2.8 | - |
D-fructose 6-phosphate | pH 6.5, 40°C, mutant E72A | uncultured bacterium |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Pyrococcus horikoshii | |
Mg2+ | required | Methanocaldococcus jannaschii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + D-fructose 6-phosphate | uncultured bacterium | - |
AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | Pyrococcus horikoshii | - |
AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | Methanocaldococcus jannaschii | - |
AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | Methanocaldococcus jannaschii ATCC 43067 | - |
AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | Pyrococcus horikoshii ATCC 700860 | - |
AMP + D-fructose 1,6-bisphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | Q58999 | - |
- |
Pyrococcus horikoshii | O59355 | - |
- |
Pyrococcus horikoshii ATCC 700860 | O59355 | - |
- |
uncultured bacterium | - |
synthetic construct | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography | uncultured bacterium |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography | Pyrococcus horikoshii |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography | Methanocaldococcus jannaschii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + D-fructose 6-phosphate | - |
uncultured bacterium | AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | - |
Pyrococcus horikoshii | AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | - |
Methanocaldococcus jannaschii | AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | - |
Methanocaldococcus jannaschii ATCC 43067 | AMP + D-fructose 1,6-bisphosphate | - |
? | |
ADP + D-fructose 6-phosphate | - |
Pyrococcus horikoshii ATCC 700860 | AMP + D-fructose 1,6-bisphosphate | - |
? | |
additional information | the enzyme also shows glucokinase activity, a bifunctional AncPFK/GK ancestor enzyme | uncultured bacterium | ? | - |
- |
|
additional information | the enzyme from Methanococcus jannaschii also shows glucokinase activity, a bifunctional MjPFK/GK | Methanocaldococcus jannaschii | ? | - |
- |
|
additional information | the PFK from Pyrococcus horikoshii (PhPFK) has no activity with glucose | Pyrococcus horikoshii | ? | - |
- |
|
additional information | the enzyme from Methanococcus jannaschii also shows glucokinase activity, a bifunctional MjPFK/GK | Methanocaldococcus jannaschii ATCC 43067 | ? | - |
- |
|
additional information | the PFK from Pyrococcus horikoshii (PhPFK) has no activity with glucose | Pyrococcus horikoshii ATCC 700860 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
ADP-dependent glucokinase | - |
uncultured bacterium |
ADP-GK | - |
uncultured bacterium |
ancGK/PFK | - |
uncultured bacterium |
MjPFK/GK | - |
Methanocaldococcus jannaschii |
More | see also EC 2.7.1.147 | uncultured bacterium |
More | see also EC 2.7.1.147 | Methanocaldococcus jannaschii |
PFK | - |
Pyrococcus horikoshii |
PhPFK | - |
Pyrococcus horikoshii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at | uncultured bacterium |
40 | - |
assay at | Pyrococcus horikoshii |
40 | - |
assay at | Methanocaldococcus jannaschii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.33 | - |
D-fructose 6-phosphate | pH 6.5, 40°C, wild-type enzyme | uncultured bacterium | |
0.38 | - |
D-fructose 6-phosphate | pH 6.5, 40°C, mutant E72A | uncultured bacterium | |
2.7 | - |
D-fructose 6-phosphate | pH 6.5, 40°C | Methanocaldococcus jannaschii | |
21 | - |
D-fructose 6-phosphate | pH 6.5, 40°C | Pyrococcus horikoshii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
assay at | uncultured bacterium |
6.5 | - |
assay at | Pyrococcus horikoshii |
6.5 | - |
assay at | Methanocaldococcus jannaschii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ADP | - |
Pyrococcus horikoshii | |
ADP | - |
Methanocaldococcus jannaschii |
General Information | Comment | Organism |
---|---|---|
evolution | consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction | uncultured bacterium |
evolution | consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction | Pyrococcus horikoshii |
evolution | consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction | Methanocaldococcus jannaschii |
additional information | molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics | uncultured bacterium |
additional information | molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics | Pyrococcus horikoshii |
additional information | molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics | Methanocaldococcus jannaschii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.136 | - |
D-fructose 6-phosphate | pH 6.5, 40°C, mutant E72A | uncultured bacterium | |
2.77 | - |
D-fructose 6-phosphate | pH 6.5, 40°C, wild-type enzyme | uncultured bacterium | |
375 | - |
D-fructose 6-phosphate | pH 6.5, 40°C | Pyrococcus horikoshii | |
428.6 | - |
D-fructose 6-phosphate | pH 6.5, 40°C | Methanocaldococcus jannaschii |