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 | 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 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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Trypanosoma brucei | 5829 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Trypanosoma brucei |
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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei | Q52MQ5 | stock 427, bifunctional enzyme, 4 isozymes | - |
Trypanosoma brucei | Q6PY95 | stock 427 | - |
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 | 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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Trypanosoma brucei |
0.011 | - |
purified enzyme | Trypanosoma brucei |
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 | 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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Trypanosoma brucei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
about | Trypanosoma brucei |
7.1 | - |
assay at | Trypanosoma brucei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Trypanosoma brucei | |
ATP | binding site of isozyme Tb1, overview | Trypanosoma brucei |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.7 | - |
phosphoenolpyruvate | 30°C | Trypanosoma brucei |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Trypanosoma brucei | isozymes, sequence calculation | - |
9.3 |