Cloned (Comment) | Organism |
---|---|
subcloning and overexpression of 2 brain enzyme variants HBP1 and HBP2 and of mutant enzymes as His6-tagged proteins in Escherichia coli, expression of enzyme form HBP1 in Spodoptera frugiperda Sf9 insect cells via baculovirus infection system | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
H253A | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
H253A/S460D | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
additional information | construction of a chimeric mutant fusing the N-terminal portion of residues 1-444 of the human enzyme to the C-terminal part, residues 450-530, of the Bos taurus enzyme, i.e. HBPBHP, construction of a deletion mutant lacking the entire carboxyterminal region of residues 446-519 | Homo sapiens |
S302R | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
S460D | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
S460D/T470D | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
S477D | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
T470D | site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
beta-D-fructose 6-phosphate | modeling of beta-D-fructose 6-phosphate as inhibitor | Homo sapiens | |
phosphoenolpyruvate | HBP1 and HBP2, the bifunctional enzyme is regulated via inhibition by phosphoenolpyruvate, uncompetitive against ATP, noncompetitive against beta-D-fructose 6-phosphate | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | recombinant wild-type and mutant enzymes, Km for phosphorylated wild-type enzyme and mutants with beta-D-fructose 6-phosphate and beta-D-fructose 2,6-bisphosphate, kinetics, modeling | Homo sapiens | |
0.0064 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant S477D | Homo sapiens | |
0.0066 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutants T470D and S460D/T470D | Homo sapiens | |
0.0067 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant S460D | Homo sapiens | |
0.0071 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant H253A/S460D | Homo sapiens | |
0.0071 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, wild-type HBP1 | Homo sapiens | |
0.0073 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant H253A | Homo sapiens | |
0.0074 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged wild-type HBP2 | Homo sapiens | |
0.008 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged wild-type HBP1 | Homo sapiens | |
0.0092 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant Hnon-phosphorylated, is-tagged HBP1 deletion mutant | Homo sapiens | |
0.0095 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged HBP1 mutant S302R | Homo sapiens | |
0.013 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged chimeric mutant enzyme, phosphorylated or nonphosphorylated | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + beta-D-fructose 6-phosphate | Homo sapiens | - |
ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
additional information | Homo sapiens | the bifunctional enzyme catalyzes the synthesis, 6-phosphofructo-2-kinase, and hydrolysis, fructose-2,6-bisphosphatase, of beta-D-fructose 2,6-bisphosphate, an activator of glycolysis and an inhibitor of gluconeogenesis | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q16875 | exon 15a and complete sequence; 10 splicing variants | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | recombinant His-tagged HBP2, and nontagged wild-type and mutant S460D HBP1 are phosphorylated by several protein kinases, i.e. protein kinases A, B, and C, and by AMP activated kinase, HBP1 mutants are phosphorylated by protein kinase C, overview | Homo sapiens |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme variants HBP1 and HBP2 and of mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and ion exchange chromatography, the tag is cleaved off, recombinant enzyme form HBP1 partially from Sf9 insect cells by affinity chromatography | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + beta-D-fructose 6-phosphate = ADP + beta-D-fructose 2,6-bisphosphate | HBP1 and HBP2 both show a random order equilibrium bi bi mechanism, bifunctional enzyme comprises both 6-phosphofructo-2-kinase and fructose-2,6-bisphosphatase, EC 3.1.3.46, activities | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
brain | - |
Homo sapiens | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.2 | - |
recombinant purified HBP1 from Escherichia coli | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + beta-D-fructose 6-phosphate | - |
Homo sapiens | 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 | Homo sapiens | ADP + beta-D-fructose 2,6-bisphosphate | - |
? | |
additional information | the bifunctional enzyme catalyzes the synthesis, 6-phosphofructo-2-kinase, and hydrolysis, fructose-2,6-bisphosphatase, of beta-D-fructose 2,6-bisphosphate, an activator of glycolysis and an inhibitor of gluconeogenesis | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase | - |
Homo sapiens |
6PF2K/Fru-2,6-P2ase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kcat of wild-type and mutant HBP1 and of HBP2 with beta-D-fructose 2,6-bisphosphate, kcat for phosphorylated wild-type enzyme and mutants with beta-D-fructose 6-phosphate and beta-D-fructose 2,6-bisphosphate | Homo sapiens | |
0.391 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant H253A | Homo sapiens | |
0.394 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant S460D | Homo sapiens | |
0.394 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant S460D/T470D | Homo sapiens | |
0.399 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged HBP1 mutant S302R | Homo sapiens | |
0.401 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant H253A/S460D | Homo sapiens | |
0.405 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant S477D | Homo sapiens | |
0.407 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant wild-type HBP1 | Homo sapiens | |
0.43 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged wild-type HBP1 | Homo sapiens | |
0.435 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant T470 | Homo sapiens | |
0.447 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged wild-type HBP2 | Homo sapiens | |
0.654 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged chimeric mutant enzyme | Homo sapiens | |
0.678 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant phosphorylated, His-tagged chimeric mutant enzyme | Homo sapiens | |
0.696 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged HBP1 deletion mutant | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | 7.5 | assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | modeling of inhibition of HBP1 and HBP2 by phosphoenolpyruvate and beta-D-fructose 6-phosphate, Km for phosphorylated wild-type enzyme and mutants with beta-D-fructose 6-phosphate and beta-D-fructose 2,6-bisphosphate | Homo sapiens | |
8.5 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant S460D/T470D | Homo sapiens | |
9.2 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant S460D | Homo sapiens | |
9.5 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant H253A/S460D | Homo sapiens | |
9.6 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged HBP1 deletion mutant | Homo sapiens | |
9.7 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant S477D | Homo sapiens | |
9.9 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant wild-type HBP1 | Homo sapiens | |
10.9 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged HBP1 mutant S302R | Homo sapiens | |
11.1 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant H253A | Homo sapiens | |
11.8 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged wild-type HBP2 | Homo sapiens | |
12.2 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant His-tagged wild-type HBP1 | Homo sapiens | |
12.7 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged chimeric mutant enzyme | Homo sapiens | |
13 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant HBP1 mutant T470 | Homo sapiens | |
13.4 | - |
beta-D-fructose 6-phosphate | pH 7.5, 30°C, recombinant phosphorylated, His-tagged chimeric mutant enzyme | Homo sapiens |