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Literature summary for 2.7.1.105 extracted from
- 6-Phosphofructo-2-kinase and fructose-2,6-bisphosphatase in Trypanosomatidae. Molecular characterization, database searches, modelling studies and evolutionary analysis (2005), FEBS J., 272, 3542-3560.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA and amino acid sequence determination and analysis | Trypanosoma brucei |
| DNA and amino acid sequence determination and analysis, phylogenetic analysis of the 6-phosphofructo-2-kinase and the fructose-2,6-bisphosphatase domains, expression of His-tagged wild-type and mutant isozymes Tb1, Tb2, and Tb4 in Escherichia coli, poor expression levels and mostly inactive and unstable isozymes, e.g. recombinant Tb2 is inactive | Trypanosoma brucei |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| citrate | 60% inhibition at 1 mM | Trypanosoma brucei |  |
| glycerol 3-phosphate | 20% inhibition at 2mM | Trypanosoma brucei | |
| additional information | enzyme is not affected by protein kinase C | Trypanosoma brucei | |
| phosphoenolpyruvate | - |
Trypanosoma brucei | |
| protein kinase A | inactivation via a 7fold increase in Km for fructose 6-phosphate without alteration of Vmax | Trypanosoma brucei |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.88 | - |
ATP | 30°C | Trypanosoma brucei | |
| 1.62 | - |
ATP | pH 7.1, 30°C, recombinant isozyme Tb1 | Trypanosoma brucei | |
| 1.9 | 4.6 | beta-D-fructose 6-phosphate | pH 7.1, 30°C, recombinant isozyme Tb1 | Trypanosoma brucei | |
| 5.8 | - |
beta-D-fructose 6-phosphate | pH 7.1, 30°C | Trypanosoma brucei | |
| 39 | - |
beta-D-fructose 6-phosphate | pH 7.1, 30°C, protein kinase A treated enzyme | Trypanosoma brucei | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| cytosol | - |
Trypanosoma brucei | 5829 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Trypanosoma brucei | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 54000 | - |
x * 72000, isozyme Tb2, about, sequence calculation, x * 54000, isozyme Tb3, about, sequence calculation, x * 79000, isozyme Tb4, about, sequence calculation | Trypanosoma brucei |
| 60000 | - |
2 * 60000, recombinant Tb1, SDS-PAGE, 2 * 111000, isozyme Tb1, about, sequence calculation | Trypanosoma brucei |
| 72000 | - |
x * 72000, isozyme Tb2, about, sequence calculation, x * 54000, isozyme Tb3, about, sequence calculation, x * 79000, isozyme Tb4, about, sequence calculation | Trypanosoma brucei |
| 76400 | - |
gel filtration | Trypanosoma brucei |
| 79000 | - |
x * 72000, isozyme Tb2, about, sequence calculation, x * 54000, isozyme Tb3, about, sequence calculation, x * 79000, isozyme Tb4, about, sequence calculation | Trypanosoma brucei |
| 111000 | - |
2 * 60000, recombinant Tb1, SDS-PAGE, 2 * 111000, isozyme Tb1, about, sequence calculation | Trypanosoma brucei |
| 140000 | - |
recombinant isozyme Tb1, gel filtration | Trypanosoma brucei |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + beta-D-fructose 6-phosphate | Trypanosoma brucei | - |
ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
| additional information | Trypanosoma brucei | evolution of the bifunctional enzyme | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Trypanosoma brucei | Q52MQ5 | stock 427, bifunctional enzyme, 4 isozymes | - |
| Trypanosoma brucei | Q6PY95 | stock 427 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| from cytosol, 9000fold by ion exchange and affinity chromatography | Trypanosoma brucei |
| native enzyme from stock 427 by ion exchange and affinity chromatography, recombinant His-tagged wild-type and mutant isozyme Tb1 from Escherichia coli | Trypanosoma brucei |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| ATP + beta-D-fructose 6-phosphate = ADP + beta-D-fructose 2,6-bisphosphate | bifunctional enzyme comprises both 6-phosphofructo-2-kinase and fructose-2,6-bisphosphatase, EC 3.1.3.46, activities, residues K51, T52, D128, and K172 are key catalytic residues for the 6-phosphofructo-2-kinase activity | Trypanosoma brucei |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Trypanosoma brucei |
| 0.011 | - |
purified enzyme | Trypanosoma brucei |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + beta-D-fructose 6-phosphate | - |
Trypanosoma brucei | ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
| ATP + beta-D-fructose 6-phosphate | bifunctional enzyme catalyzes the forward and reverse reaction using different catalytic sites | Trypanosoma brucei | ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
| additional information | evolution of the bifunctional enzyme | Trypanosoma brucei | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 72000, isozyme Tb2, about, sequence calculation, x * 54000, isozyme Tb3, about, sequence calculation, x * 79000, isozyme Tb4, about, sequence calculation | Trypanosoma brucei |
| dimer | 2 * 60000, recombinant Tb1, SDS-PAGE, 2 * 111000, isozyme Tb1, about, sequence calculation | Trypanosoma brucei |
| More | bifunctional enzyme domain structure, the bifunctional enzyme possesses a 6-phosphofructo-2-kinase and a fructose-2,6-bisphosphatase domain, as well as ankyrin-motif repeats, overview | Trypanosoma brucei |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase | - |
Trypanosoma brucei |
| Pfk-2 | - |
Trypanosoma brucei |
| PFK-2/FBPase-2 | - |
Trypanosoma brucei |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Trypanosoma brucei |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
about | Trypanosoma brucei |
| 7.1 | - |
assay at | Trypanosoma brucei |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Trypanosoma brucei | |
| ATP | binding site of isozyme Tb1, overview | Trypanosoma brucei | |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.7 | - |
phosphoenolpyruvate | 30°C | Trypanosoma brucei | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Trypanosoma brucei | isozymes, sequence calculation | - |
9.3 |
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