Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Manes, N.P.; El-Maghrabi, M.R.
    The kinase activity of human brain 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase is regulated via inhibition by phosphoenolpyruvate (2005), Arch. Biochem. Biophys., 438, 125-136.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.105 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

EC Number Protein Variants Comment Organism
2.7.1.105 H253A site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens
2.7.1.105 H253A/S460D site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens
2.7.1.105 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
2.7.1.105 S302R site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens
2.7.1.105 S460D site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens
2.7.1.105 S460D/T470D site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens
2.7.1.105 S477D site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens
2.7.1.105 T470D site-directed mutagenesis, altered kinetics compared to wild-type enzyme, overview Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.105 beta-D-fructose 6-phosphate modeling of beta-D-fructose 6-phosphate as inhibitor Homo sapiens
2.7.1.105 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.105 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
2.7.1.105 0.0064
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant S477D Homo sapiens
2.7.1.105 0.0066
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutants T470D and S460D/T470D Homo sapiens
2.7.1.105 0.0067
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant S460D Homo sapiens
2.7.1.105 0.0071
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant H253A/S460D Homo sapiens
2.7.1.105 0.0071
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, wild-type HBP1 Homo sapiens
2.7.1.105 0.0073
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, HBP1 mutant H253A Homo sapiens
2.7.1.105 0.0074
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged wild-type HBP2 Homo sapiens
2.7.1.105 0.008
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged wild-type HBP1 Homo sapiens
2.7.1.105 0.0092
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant Hnon-phosphorylated, is-tagged HBP1 deletion mutant Homo sapiens
2.7.1.105 0.0095
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged HBP1 mutant S302R Homo sapiens
2.7.1.105 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

EC Number Metals/Ions Comment Organism Structure
2.7.1.105 Mg2+
-
 Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.105 ATP + beta-D-fructose 6-phosphate Homo sapiens
-
 ADP + beta-D-fructose 2,6-bisphosphate
-
 ?
2.7.1.105 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

EC Number Organism UniProt Comment Textmining
2.7.1.105 Homo sapiens Q16875 exon 15a and complete sequence; 10 splicing variants
-

Posttranslational Modification

EC Number Posttranslational Modification Comment Organism
2.7.1.105 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 (Commentary)

EC Number Purification (Comment) Organism
2.7.1.105 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

EC Number Reaction Comment Organism Reaction ID
2.7.1.105 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
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.105 brain
-
 Homo sapiens
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.105 0.2
-
 recombinant purified HBP1 from Escherichia coli Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.105 ATP + beta-D-fructose 6-phosphate
-
 Homo sapiens ADP + beta-D-fructose 2,6-bisphosphate
-
 ?
2.7.1.105 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
-
 ?
2.7.1.105 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

EC Number Synonyms Comment Organism
2.7.1.105 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase
-
 Homo sapiens
2.7.1.105 6PF2K/Fru-2,6-P2ase
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.105 30
-
 assay at Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.105 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
2.7.1.105 0.391
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant H253A Homo sapiens
2.7.1.105 0.394
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant S460D Homo sapiens
2.7.1.105 0.394
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant S460D/T470D Homo sapiens
2.7.1.105 0.399
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged HBP1 mutant S302R Homo sapiens
2.7.1.105 0.401
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant H253A/S460D Homo sapiens
2.7.1.105 0.405
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant S477D Homo sapiens
2.7.1.105 0.407
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant wild-type HBP1 Homo sapiens
2.7.1.105 0.43
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged wild-type HBP1 Homo sapiens
2.7.1.105 0.435
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant T470 Homo sapiens
2.7.1.105 0.447
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged wild-type HBP2 Homo sapiens
2.7.1.105 0.654
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged chimeric mutant enzyme Homo sapiens
2.7.1.105 0.678
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant phosphorylated, His-tagged chimeric mutant enzyme Homo sapiens
2.7.1.105 0.696
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged HBP1 deletion mutant Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.105 7 7.5 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.105 ATP
-
 Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.7.1.105 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
2.7.1.105 8.5
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant S460D/T470D Homo sapiens
2.7.1.105 9.2
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant S460D Homo sapiens
2.7.1.105 9.5
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant H253A/S460D Homo sapiens
2.7.1.105 9.6
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged HBP1 deletion mutant Homo sapiens
2.7.1.105 9.7
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant S477D Homo sapiens
2.7.1.105 9.9
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant wild-type HBP1 Homo sapiens
2.7.1.105 10.9
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged HBP1 mutant S302R Homo sapiens
2.7.1.105 11.1
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant H253A Homo sapiens
2.7.1.105 11.8
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged wild-type HBP2 Homo sapiens
2.7.1.105 12.2
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant His-tagged wild-type HBP1 Homo sapiens
2.7.1.105 12.7
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant non-phosphorylated, His-tagged chimeric mutant enzyme Homo sapiens
2.7.1.105 13
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant HBP1 mutant T470 Homo sapiens
2.7.1.105 13.4
-
 beta-D-fructose 6-phosphate pH 7.5, 30°C, recombinant phosphorylated, His-tagged chimeric mutant enzyme Homo sapiens