Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
L307A | site-directed mutagenesis, C-terminal point mutant, further mutants analyzed that reveal successive deletions of up to 10 residues at the C-terminal end | Escherichia coli |
Y306A | site-directed mutagenesis, C-terminal point mutant, no dimer-tetramer conversion in presence of MgATP, inhibition pattern almost undistinguishable from the wild-type, conformational changes leading to allosteric inhibition can be uncoupled from tetramer formation at least in the Y306A mutant | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
MgATP2- | allosteric inhibition | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
MgATP2- | C-terminal deletion mutant L307, 1 mM co-substrate | Escherichia coli | |
0.0137 | - |
MgATP2- | Y306A, C-terminal point mutant, 1 mM co-substrate | Escherichia coli | |
0.015 | - |
MgATP2- | wild-type, 1 mM co-substrate | Escherichia coli | |
0.02 | - |
beta-D-fructose 6-phosphate | C-terminal deletion mutant D299, 1 mM co-substrate | Escherichia coli | |
0.029 | - |
beta-D-fructose 6-phosphate | L307A, C-terminal point mutant, 1 mM co-substrate | Escherichia coli | |
0.036 | - |
beta-D-fructose 6-phosphate | C-terminal deletion mutant A305, 1 mM co-substrate | Escherichia coli | |
0.057 | - |
beta-D-fructose 6-phosphate | wild-type, 1 mM co-substrate, chosen in order avoid the effect of MgATP binding at the allosteric site to the binding of fructose-6-P to the catalytic site, that is observed above 1 mM MgATP | Escherichia coli | |
0.06 | - |
beta-D-fructose 6-phosphate | C-terminal deletion mutant L307, 1 mM co-substrate | Escherichia coli | |
0.062 | - |
beta-D-fructose 6-phosphate | Y306A, C-terminal point mutant, 1 mM co-substrate | Escherichia coli | |
0.8 | - |
MgATP2- | C-terminal deletion mutant A305, 1 mM co-substrate | Escherichia coli | |
0.88 | - |
MgATP2- | C-terminal deletion mutant Y306, 1 mM co-substrate | Escherichia coli | |
0.954 | - |
MgATP2- | L307A, C-terminal point mutant, 1 mM co-substrate | Escherichia coli | |
2 | - |
MgATP2- | C-terminal deletion mutant D299, 1 mM co-substrate | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required for activity | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + beta-D-fructose 6-phosphate | Escherichia coli | - |
ADP + beta-D-fructose 2,6-bisphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
gel filtration | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
binding of MgATP to an allosteric site provokes inhibition and a dimer-tetramer conversion of the enzyme, mutant generation by successive deletions of up to 10 residues and point mutations at the C-terminal end, elevated KM values for MgATP that fails to show the dimer-tetramer conversion, Y306 required for the quaternary packing involved in the dimer-tetramer conversion, the following residue L307 is crucial for the ternary packing necessary for the catalytic MgATP-binding site, dimer-tetramer conversion can be uncoupled from the conformational changes that lead to the MgATP-induced allosteric inhibition | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + beta-D-fructose 6-phosphate | - |
Escherichia coli | ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
ATP + beta-D-fructose 6-phosphate | activity determined spectrophotometrically by coupling the fructose-1,6-bisphosphate production to the oxidation of NADH | Escherichia coli | ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
MgATP2- + beta-D-fructose 6-phosphate | analysis of MgATP-induced tetramer formation | Escherichia coli | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Pfk-2 | - |
Escherichia coli |