Information on EC 3.1.3.46 - fructose-2,6-bisphosphate 2-phosphatase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.1.3.46
-
RECOMMENDED NAME
GeneOntology No.
fructose-2,6-bisphosphate 2-phosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
beta-D-fructose 2,6-bisphosphate + H2O = D-fructose 6-phosphate + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
fructose 2,6-bisphosphate biosynthesis
-
-
Fructose and mannose metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
beta-D-fructose-2,6-bisphosphate 2-phosphohydrolase
The enzyme copurifies with EC 2.7.1.105 6-phosphofructo-2-kinase. (cf. EC 3.1.3.54 fructose-2,6-bisphosphate 6-phosphatase).
CAS REGISTRY NUMBER
COMMENTARY hide
81611-75-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase long form; gene Pfrx
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase
-
-
Manually annotated by BRENDA team
Mus musculus C57/BL6
C57/BL6
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
wild type, two mutants with suppressed expression of F2KP, Tos 17 insertion mutant of OsF2KP1, RNAi mutants of OsF2KP2
-
-
Manually annotated by BRENDA team
Pigeon
-
-
-
Manually annotated by BRENDA team
Wistar
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
stock 427, isozymes 1-4
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
knockdown of isoform PFKFB4 reduces tumor growth, glucose uptake and beta-D-fructose 2,6-bisphosphate and increases apoptosis
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
beta-D-fructose 2,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
show the reaction diagram
D-fructose 2,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
show the reaction diagram
fructose 2,6-bisphosphate + H2O
?
show the reaction diagram
fructose 2,6-bisphosphate + H2O
fructose 6-phosphate + phosphate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
beta-D-fructose 2,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
show the reaction diagram
D-fructose 2,6-bisphosphate + H2O
D-fructose 6-phosphate + phosphate
show the reaction diagram
-
-
-
-
?
fructose 2,6-bisphosphate + H2O
?
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
can replace Mg2+
Mn2+
-
can replace Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2E)-2-(1,3-benzodioxol-5-ylmethylidene)butanedioic acid
-
mimics binding pattern of fructose 6-phosphate
([2-[(5-nitropyridin-2-yl)amino]phenyl]sulfanyl)acetic acid
-
mimics binding pattern of fructose 6-phosphate
1,1'-ethane-1,2-diylbis(4-acetylpyrrolidine-2,3,5-trione)
-
mimics binding pattern of fructose 6-phosphate
1-(2,3-dihydro-1,4-benzodioxin-6-yl)-2-(2-methyl-4-nitro-1H-imidazol-1-yl)ethanol
-
mimics binding pattern of fructose 6-phosphate
1-amino-4-(2-nitro-1H-imidazol-1-yl)butan-2-ol
-
mimics binding pattern of fructose 6-phosphate
2,5-anhydro-D-mannitol 6-phosphate
-
-
2-((5-bromo-6-oxo-1-phenyl-1,6-dihydropyridazin-4-yl)amino)acetamide
-
-
-
2-(1,3-benzodioxol-5-ylmethyl)-3-methylbutanedioic acid
-
mimics binding pattern of fructose 6-phosphate
2-(2-nitrophenoxy)-N-phenylacetamide
-
mimics binding pattern of fructose 6-phosphate
2-(3H-indol-7-ylmethyl)butanedioic acid
-
mimics binding pattern of fructose 6-phosphate
2-(5-amino-4-carbamoyl-1H-pyrazol-1-yl)ethanesulfonic acid
-
mimics binding pattern of fructose 6-phosphate
2-(5-bromo-6-oxo-1-phenyl-1,6-dihydropyridazin-4-yl)-1,2,3,4-tetrahydroisoquinoline-5-carbonitrile
-
-
-
2-(acetylamino)-beta-oxophenylalanine
-
mimics binding pattern of fructose 6-phosphate
2-nitro-N-(pyrazin-2-ylmethyl)benzenesulfonamide
-
mimics binding pattern of fructose 6-phosphate
2-nitro-N-quinolin-3-ylbenzenesulfonamide
-
mimics binding pattern of fructose 6-phosphate
2-[(4-nitro-1H-benzimidazol-7-yl)sulfanyl]ethyl benzoate
-
mimics binding pattern of fructose 6-phosphate
2-[(5-nitropyridin-2-yl)amino]ethyl 5-nitro-1H-pyrrole-2-carboxylate
-
mimics binding pattern of fructose 6-phosphate
2-[(furan-2-ylmethyl)amino]-5-nitrobenzoic acid
-
mimics binding pattern of fructose 6-phosphate
2-[[(2Z)-2-(phenylhydrazono)acetyl]amino]benzoic acid
-
mimics binding pattern of fructose 6-phosphate
2-[[2-(2,4-dinitrophenyl)hydrazino]carbonyl]benzoic acid
-
mimics binding pattern of fructose 6-phosphate
3,3'-(2,4,6-trioxo-1,3,5-triazinane-1,3-diyl)dipropanoic acid
-
mimics binding pattern of fructose 6-phosphate
3-(3-pyridin-2-yl-1,2,4-oxadiazol-5-yl)-N-(tetrahydrofuran-2-ylmethyl)propanamide
-
mimics binding pattern of fructose 6-phosphate
3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one
-
-
3-acetylphenyl (3-acetylphenyl)acetate
-
mimics binding pattern of fructose 6-phosphate
3-[(2-pyridin-2-ylhydrazino)carbonyl]pyrazine-2-carboxylic acid
-
mimics binding pattern of fructose 6-phosphate
4-(4-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-5-bromo-6-oxopyridazin-1(6H)-yl)benzonitrile
-
-
-
4-bromo-2-phenyl-5-(((tetrahydrofuran-2-yl)methyl)amino)pyridazin-3(2H)-one
-
-
-
4-bromo-2-phenyl-5-(2-oxa-6-azaspiro[3.3]heptan-6-yl)pyridazin-3(2H)-one
-
-
-
4-bromo-5-morpholino-2-phenylpyridazin-3(2H)-one
-
-
-
5,6,7,8-tetrahydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one
-
competitive inhibitor
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-2-benzyl-4-bromopyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-2-benzylpyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(3-phenylpropyl)pyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-((2-(dimethylamino)ethyl)-amino)benzyl)pyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-(trifluoromethoxy)phenyl)-pyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-chlorophenyl)pyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-iodobenzyl)pyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(pyrimidin-5-yl)pyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-phenethylpyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-phenylpyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-chloro-2-phenylpyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-ethoxy-2-phenylpyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-iodo-2-phenylpyridazin-3(2H)-one
-
-
-
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-isopropyl-2-phenylpyridazin-3(2H)-one
-
-
-
5-hydroxy-N-(4-nitro-1,3-thiazol-2-yl)-2,4-dioxopentanamide
-
mimics binding pattern of fructose 6-phosphate
6-phospho-gluconate
-
-
6-phosphogluconate
-
citrate
-
inhibits the cardiac enzyme
Cyclic AMP dependent protein kinase
-
causes inactivation
-
D-fructose 6-phosphate
D-fructose-1,6-bisphosphate
-
D-fructose-6-phosphate
-
D-glucose-1,6-bisphosphate
-
D-psicose 6-phosphate
-
poor inhibitor
D-ribose 5-phosphate
-
poor inhibitor
D-tagatose 6-phosphate
-
poor inhibitor
diethyldicarbonate
-
inactivation
dimethyloxalylglycine
inhibits the splice isozyme PFKFB-4
ethyl 1-(6-oxo-1-phenyl-5-(2-oxa-6-azaspiro[3.3]heptan-6-yl)-1,6-dihydropyridazin-4-yl)-1H-1,2,3-triazole-4-carboxylate
-
-
-
F-
-
in presence of glycerol 3-or 2-phosphate
fructose 6-phosphate
glycerol 1-phosphate
Insulin
-
opposes action of glucagon or epinephrine
-
L-Sorbose 6-phosphate
-
-
N-Bromoacetylethanolamine
p-mercuribenzoate
-
-
phosphate
phosphoenolpyruvate
-
-
Tolbutamide
-
-
vanadate
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-phosphoglycerate
-
-
brazilin
-
a component isolated from Caesalpinia sappan wood, increases enzyme activity by increasing the cellular content of fructose-2,6-bishosphate via increased levels of fructose 6-phosphate and hexose 6-phosphate produced by activates 6-phosphofructo-2-kinase and pyruvate kinase, overview
citrate
-
-
Cyclic AMP dependent protein kinase
dihydroxyacetone phosphate
-
-
epinephrine
-
-
formate
-
activates, interacts with Glu327
glycerol 1-phosphate
Glycerol 2-phosphate
MgATP2-
-
-
MgGTP2-
phosphate
phosphoenolpyruvate
-
-
Protein kinase C
Protein phosphatase 2A
-
cyclic AMP dependent protein kinase inactivation reversible by
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0102 - 0.204
beta-D-fructose 2,6-bisphosphate
0.00013
D-fructose 2,6-bisphosphate
-
pH 7.4
0.000028
D-fructose-1,6-bisphosphate
pH 6.0, 25C
0.0001 - 1.4
fructose 2,6-bisphosphate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
fructose 2,6-bisphosphate
Rattus norvegicus
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08
6-phosphogluconate
-
9.7
D-fructose 6-phosphate
-
pH 7.4
0.08
D-fructose-1,6-bisphosphate
-
0.1
D-fructose-6-phosphate
-
0.85
D-glucose-1,6-bisphosphate
-
0.15
vanadate
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1
2-((5-bromo-6-oxo-1-phenyl-1,6-dihydropyridazin-4-yl)amino)acetamide
Homo sapiens
-
IC50 above 1.0 mM, in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.5
2-(5-bromo-6-oxo-1-phenyl-1,6-dihydropyridazin-4-yl)-1,2,3,4-tetrahydroisoquinoline-5-carbonitrile
Homo sapiens
-
IC50 above 0.5 mM,in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.026
3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
0.0084
4-(4-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-5-bromo-6-oxopyridazin-1(6H)-yl)benzonitrile
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
1
4-bromo-2-phenyl-5-(((tetrahydrofuran-2-yl)methyl)amino)pyridazin-3(2H)-one, 4-bromo-2-phenyl-5-(2-oxa-6-azaspiro[3.3]heptan-6-yl)pyridazin-3(2H)-one
0.0034
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-2-benzyl-4-bromopyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
10
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-2-benzylpyridazin-3(2H)-one
Homo sapiens
-
IC50 above 10 mM, in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.011
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(3-phenylpropyl)pyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.5
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-((2-(dimethylamino)ethyl)-amino)benzyl)pyridazin-3(2H)-one
Homo sapiens
-
IC50 above 0.5 mM, in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.0091
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-(trifluoromethoxy)phenyl)-pyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.007
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-chlorophenyl)pyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.0087
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(4-iodobenzyl)pyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.0096
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-(pyrimidin-5-yl)pyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.0026
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-phenethylpyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.0074
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-bromo-2-phenylpyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.0262
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-chloro-2-phenylpyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
1
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-ethoxy-2-phenylpyridazin-3(2H)-one
Homo sapiens
-
IC50 above 1.0 mM, in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.013
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-iodo-2-phenylpyridazin-3(2H)-one
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.5
5-(4-acetyl-5-methyl-1H-1,2,3-triazol-1-yl)-4-isopropyl-2-phenylpyridazin-3(2H)-one
Homo sapiens
-
IC50 above 0.5 mM, in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
0.055
ethyl 1-(6-oxo-1-phenyl-5-(2-oxa-6-azaspiro[3.3]heptan-6-yl)-1,6-dihydropyridazin-4-yl)-1H-1,2,3-triazole-4-carboxylate
Homo sapiens
-
in 40 mM Tris pH 7.5, 20 mM MgCl2, 0.1 mg/ml bovine serum albumin, at 30C
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000011
-
hepatocyte
0.000014
-
hepatocyte treated with brazilin
20 - 40
1650
phosphatase activity
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at
8
-
assay at
additional information
-
pH-dependence of kinetic parameters
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9.29
-
amino acid sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
adipocyte cell line
Manually annotated by BRENDA team
-
overexpression of isozyme PFKFB-4 in breast and colon malignant tumors compared to non-malignant tissue, overexpression of isozyme PFKFB-4 is correlated with enhanced expression of isozyme PFKFB-3, hypoxia-inducible factor 1alpha, dependent glucose transporter 1, and vascular endothelial growth factor VEGF, overview
Manually annotated by BRENDA team
-
overexpression of isozyme PFKFB-4 in breast and colon malignant tumors compared to non-malignant tissue, overexpression of isozyme PFKFB-4 is correlated with enhanced expression of isozyme PFKFB-3, hypoxia-inducible factor 1alpha, dependent glucose transporter 1, and vascular endothelial growth factor VEGF, overview
Manually annotated by BRENDA team
melanoma cell line, minor splice isozyme PFKFB-4, major isozyme is PFKFB-3, isozyme PFKFB-4 is overexpressed in hypoxic conditions
Manually annotated by BRENDA team
-
epididymal, reduced expression of isozyme PFKFB3
Manually annotated by BRENDA team
minor splice isozyme PFKFB-4, major isozyme is PFKFB-3
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
bloodstream form, grown in rats
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
12000
-
gel filtration
76200
-
gel filtration
100000
107000
-
equilibrium sedimentation
110000
140000
-
isozyme 2, gel filtration
320000
gel filtration
370000
gel filtration
600000
-
about, isozyme 1, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
-
1* 36600 and 1* 35600 which seems a degradation product of the larger subunit, SDS-PAGE
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
as hanging drops in vapor diffusion VDX plates with Mg2+, citrate and beta-D-fructose 2,6-bisphosphate
-
by the sitting-drop, vapor-diffusion method, crystals of PFKFB D-fructose 6-phosphate complex in the presence of the nonreactive ATP-analogue AMPPCP beta,gamma-methyleneadenosine 5-triphosphate and crystals the inhibitory complex of PFKFB ADP and phosphoenolpyruvate
-
crystal structure of the liver enzyme
-
in complex with D-fructose 6-phosphate, diphosphate, or AlF4, sitting drop vapor diffusion method, using 100 mM Tris-HCl, pH 7.5, 20-25% (w/v) ethylene glycol, 200-400 mM tartaric acid, 5% (v/v) glycerol, and 12% (w/v) polyethylene glycol 4000
purified recombinant inducible isozyme PFKFB3, 8 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 10 mM sodium phosphate, 0.05 mM EDTA, 5 mM 2-mercaptoethanol, 0.2 mM ADP, 5% glycerol, and 0.2 mM fructose-6-phosphate, sitting drop vapour diffusion method in presence or absence of fructose-2,6-bisphosphate, the protein sample is mixed with an equal volume of mother liquor containing 50 mM Tris-HCl, pH 7.5, 20-25% ethylene glycol, 12% dioxane, 5% glycerol, and 12% PEG 4000, 2-3 weeks, removal of free phosphate from crystals prior to X-ray diffraction structure determination and analysis at 2.1 A resolution; sitting-drop vapor diffusion of the 1:1 mixture of the protein sample with mother liquor of 50 mM Tris-HCl, pH 7.5, 20.35% ethylene glycol, 12% dioxane, 5% glycerol, and 12% polyethylene glycol 4000. Crystals in a size of 0.2 * 0.2 * 0.05 mm grow in 2-3 weeks. Determination of structure at 2.1 A resolution
as hanging drops in vapor diffusion VDX plates in complex with Mg2+ or Zn2+ and beta-D-fructose 2,6-bisphosphate
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42
10 min, inactivation, phosphate protects
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 20% glycerol
-
-70C for 3 months including 100 mM KCl, 0.5 mM dithiothreitol, 20% glycerol and 50 mM Tris-HCl at pH 7.4
-
-70C, 20 mM sodium phosphate buffer, pH 7.3, 1 mM dithiothreitol
-
-80C
-
-80C, 10% glycerol, more than 1 week
-
-80C, 20% glycerol, 200 mM KCl, 1 mM dithiothreitol, 20 mM Hepes, pH 7.1
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
10000fold
-
15fold
-
bifunctional enzyme, copurification with phosphofructokinase 2
by Cibacron Blue-Sepharose and Sephadex G-50 column chromatography
by Sephadex G-25 column chromatography
-
copurification with phosphofructokinase 2
-
glutathione Sepharose 4 column chromatography
-
Ni-NTA affinity column chromatography
partially from liver
-
recombinant His-tagged inducible isozyme PFKFB3 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, after which the tag is cleaved off by thrombin, followed by anion exchange chromatography, to homogeneity
recombinant His-tagged isozymes 1-4 from purified trypomastigotes, by anion exchange and adsorption chromatography, ultrafiltration and dialysis
-
recombinant His6-tagged bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography and anion exchange chromatography with triethylammonium bicarbonate as eluant
-
two isoenzymes
-
two isoenzymes, partially purified
-
using Ni-NTA affinity columns
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
as selenomethionine-substituted protein
-
bifunctional enzyme, isozymes 1-4, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of His-tagged isozymes 1-4 in Escherichia coli
-
DNA and amino acid sequence determination and analysis, phylogenetic analysis
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Mus musculus cardiac cells
-
expressed in RINm5F-GK cells
-
expression in Escherichia coli
expression in hepatocytes by incubation for 2h with increasing titres of adenoviral vectors
-
expression in MIN-6 cell by incubation for 2h with increasing titres of adenoviral vectors
-
expression of His-tagged inducible isozyme PFKFB3 in Escherichia coli strain BL21(DE3)
expression of His6-tagged bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in Escherichia coli strain BL21(DE3) pLysS
-
expression of wild-type and mutant PFK-2 in 6-phosphofructo-2-kinase-deficient myocytes
-
for the RNAi construct using the JJ374 vector and Agrobacterium tumefaciens for transformation
-
gene PFKFB4, DNA and amino acid sequence determination, expression analysis in DB-1 melanoma cells
genes PFKFB1-4, chromosomal localization of tissue-specific isozymes, phylogenetic analysis, overview, transcriptional control of isozymes, overview; genes PFKFB1-4, chromosomal localization of tissue-specific isozymes, phylogenetic analysis, overview, transcriptional control of isozymes, overview; genes PFKFB1-4, chromosomal localization of tissue-specific isozymes, phylogenetic analysis, overview, transcriptional control of isozymes, overview; genes PFKFB1-4, chromosomal localization of tissue-specific isozymes, phylogenetic analysis, overview, transcriptional control of isozymes, overview
isozyme PFKFB3 from 3T3-L1 adipocytes, expression and splicing analysis, overview, expression as His-tagged enzyme in Escherichia coli, expression of His-tagged enzyme in COS-7 cells
-
KDO-phosphatase knockout, enzyme is encoded by 4 genes, PFKFB1-4, PFKFB3 is activated by mitogenic, inflammatory and hypoxic stimuli
-
overexpression in Arabidopsis using the Agrobacterium tumefaciens GV3101 mediated floral dip method, gene knockout mutants and RNAi mutants, overexpression induces increased levels of soluble sugars, declined starch and triose phosphate content, beta-D-fructose 2,6-bisphosphate contributes to the regulation of starch and sucrose levels
-
overexpression in Escherichia coli as His6-tagged fusion protein
-
overexpression in hepatocytes of the wild type enzyme and of the kinase active double mutant S32A/H258A, overexpression of glukokinase and PFK have a synergistic effect on beta-D-fructose 2,6-bisphosphate levels
-
overexpression of hepatic 6-phosphofructo-2-kinase/fructose-2,6-bisphosphate using an adenoviral vector
-
overexpression of PFK-2/FBPase-2 in MIN6 cells
-
phylogenetic analysis
phylogenetic analysis, transcriptional control of isozymes, overview; phylogenetic analysis, transcriptional control of isozymes, overview
phylogenetic analysis, transcriptional control of isozymes, overview; phylogenetic analysis, transcriptional control of isozymes, overview; phylogenetic analysis, transcriptional control of isozymes, overview
possible evolution of the proteobacterial gene by horizontal gene transfer from an eukaryote, phylogenetic analysis
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
androgen stimulates glycolysis for de novo lipid synthesis by increasing the activities of hexokinase 2 and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 2, up-regulation of PFKFB2 expression in LNCaP cells is mediated by the direct binding of ligand-activated androgen receptor to the PFKFB2 promoter. Expression of PFKFB2 gene is increased 3.0fold by treatment with methyltrienolone, i.e. R1881, a synthetic androgen
-
the enzyme is induced in hypoxia and upregulated as a consequence of the transcriptional changes orchestrated by the androgen receptor in prostate cancer cells
-
the enzyme isoform PFKFB4 is about 3fold overexpressed in lung adenocarcinoma compared to normal lung tissue. PFKFB4 mRNA and protein expression are also increased by hypoxia
the protein levels are markedly elevated in high-grade astrocytomas relative to low-grade astrocytomas and corresponding non-neoplastic brain tissue, whereas no significant increase of PFKFB3 mRNA is observed in high-grade astrocytomas
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H444A/R445A/R447A
-
enhances catalytic rate of bisphosphatase activity and abrogates inhibition by kinase domain
P456E/P457A/A458E
-
change of chicken sequence to rat sequence, enzyme behaves like rat enzyme
A44G
-
site directed mutagenesis
A44V
-
site directed mutagenesis
K168A
-
site directed mutagenesis
K168N
-
site directed mutagenesis
K168R
-
site directed mutagenesis
E325A
-
role of E325 as acid-base during reaction
H258A
-
site-directed mutagenesis, mutation of the strong nucleophile His258 reduces activity compared to the wild-type enzyme
H446A
-
site-directed mutagenesis, the mutation perturbs significantly and specificially the resonances at the catalytic site assigned to His258 and His392 in His-specific multinuclear NMR study, interaction of residue 446 with R397 is disrupted, comparison to the wild-type enzyme, overview
additional information