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

-

94948

Cyclic AMP dependent protein kinase

6 entries

-

dihydroxyacetone phosphate

-

EDTA

-

epinephrine

-

formate

-

activates, interacts with Glu327

663563

glucagon

-

94934, 94939, 94941

glycerol 1-phosphate

7 entries

Glycerol 2-phosphate

GTP

-

94933, 94939

ITP

-

MgATP2-

-

MgGTP2-

phosphate

phosphoenolpyruvate

-

Protein kinase C

-

Protein phosphatase 2A

-

cyclic AMP dependent protein kinase inactivation reversible by

94931

-

salicylate

-

additional information

5 entries

-

0.0102 - 0.204

beta-D-fructose 2,6-bisphosphate

0.00013

D-fructose 2,6-bisphosphate

Gallus gallus

-

pH 7.4

650442

0.000028

D-fructose-1,6-bisphosphate

pH 6.0, 25°C

0.0001 - 1.4

fructose 2,6-bisphosphate

21 entries

additional information

-

0.1

fructose 2,6-bisphosphate

-

94933, 94945

0.08

6-phosphogluconate

-

9.7

D-fructose 6-phosphate

Gallus gallus

-

pH 7.4

650442

0.08

D-fructose-1,6-bisphosphate

-

0.1

D-fructose-6-phosphate

-

0.85

D-glucose-1,6-bisphosphate

-

0.5

phosphate

-

0.15

vanadate

Gallus gallus

-

94954

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 30°C

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 30°C

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 30°C

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 30°C

1

4-bromo-2-phenyl-5-(((tetrahydrofuran-2-yl)methyl)amino)pyridazin-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 30°C

1

4-bromo-2-phenyl-5-(2-oxa-6-azaspiro[3.3]heptan-6-yl)pyridazin-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 30°C

1

4-bromo-5-morpholino-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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

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 30°C

0.000011

-

hepatocyte

665854

0.000014

-

hepatocyte treated with brazilin

20 - 40

1650

phosphatase activity

additional information

5.5

7

7.3

-

7.5

-

assay at

713740

7.6

8

-

assay at

663563

additional information

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pH-dependence of kinetic parameters

6.5 - 8

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37

-

assay at

713740

9.29

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amino acid sequence calculation

3T3-L1 cell

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adipocyte cell line

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-

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breast carcinoma cell

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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

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-

-

-

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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

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melanoma cell line, minor splice isozyme PFKFB-4, major isozyme is PFKFB-3, isozyme PFKFB-4 is overexpressed in hypoxic conditions

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-

-

-

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epididymal, reduced expression of isozyme PFKFB3

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-

-

-

-

-

-

-

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714090, 729519, 730530

lung

lung adenocarcinoma cell

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-

-

minor splice isozyme PFKFB-4, major isozyme is PFKFB-3

-

-

-

-

-

-

-

-

-

-

-

-

bloodstream form, grown in rats

additional information

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3e9c, download, 3D-view

3e9d, download, 3D-view

3e9e, download, 3D-view

1c7z, download, 3D-view

1c80, download, 3D-view

1c81, download, 3D-view

1fbt, download, 3D-view

1tip, download, 3D-view

12000

-

gel filtration

94951

33000

Spinacia oleracea

-

2 * 33000, SDS-PAGE

94924

50000

-

2 * 50000, SDS-PAGE

722278

53000

Pigeon

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2 * 53000, SDS-PAGE

94936

53300

-

53300, SDS-PAGE

94952

53800

Bos taurus

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53800, gel electrophoresis

94949

54000

55000

-

2 * 55000, gel electrophoresis

94944

56000

-

2 * 56000

57078

-

x * 57078, amino acid sequence calculation

58000

-

x * 58000, there are 2 isoforms of 54000 and 58000 Da, SDS-PAGE

729519

76200

Spinacia oleracea

-

gel filtration

94924

83000

4 * 83000, deduced from gene sequence

90800

4 * 90800, SDS-PAGE

92000

x * 96000, phosphorylated form, x * 92000, unphosphorylated form, SDS-PAGE

653467

96000

x * 96000, phosphorylated form, x * 92000, unphosphorylated form, SDS-PAGE

653467

100000

5 entries

107000

-

equilibrium sedimentation

94935

110000

140000

-

isozyme 2, gel filtration

320000

gel filtration

370000

gel filtration

600000

-

about, isozyme 1, gel filtration

?

dimer

homodimer

monomer

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1* 36600 and 1* 35600 which seems a degradation product of the larger subunit, SDS-PAGE

94926

tetramer

additional information

25 entries

phosphoprotein

12 entries

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

-

681437

crystal structure of the liver enzyme

-

663563

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

Q16875

730905

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

P16118

665727

as hanging drops in vapor diffusion VDX plates in complex with Mg2+ or Zn2+ and beta-D-fructose 2,6-bisphosphate

-

42

10 min, inactivation, phosphate protects

-20°C, 20% glycerol

-

94931

-70°C for 3 months including 100 mM KCl, 0.5 mM dithiothreitol, 20% glycerol and 50 mM Tris-HCl at pH 7.4

-

94933

-70°C, 20 mM sodium phosphate buffer, pH 7.3, 1 mM dithiothreitol

-

-80°C

-

-80°C, 10% glycerol, more than 1 week

-

-80°C, 20% glycerol, 200 mM KCl, 1 mM dithiothreitol, 20 mM Hepes, pH 7.1

-

10000fold

-

15fold

-

300fold

-

5600fold

Pelophylax esculentus

-

94952

bifunctional enzyme, copurification with phosphofructokinase 2

by Cibacron Blue-Sepharose and Sephadex G-50 column chromatography

by Sephadex G-25 column chromatography

-

675637

copurification with phosphofructokinase 2

-

94945

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

P16118

665727

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

Spinacia oleracea

-

94929

using Ni-NTA affinity columns

-

681437

-

as selenomethionine-substituted protein

Escherichia coli

-

680855

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

-

729389, 730905

expressed in Mus musculus cardiac cells

-

690361

expressed in RINm5F-GK cells

-

721553

expression in Escherichia coli

expression in hepatocytes by incubation for 2h with increasing titres of adenoviral vectors

-

678087

expression in MIN-6 cell by incubation for 2h with increasing titres of adenoviral vectors

-

678087

expression of His-tagged inducible isozyme PFKFB3 in Escherichia coli strain BL21(DE3)

P16118

665727

expression of His6-tagged bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in Escherichia coli strain BL21(DE3) pLysS

-

713740

expression of wild-type and mutant PFK-2 in 6-phosphofructo-2-kinase-deficient myocytes

-

665582

for the RNAi construct using the JJ374 vector and Agrobacterium tumefaciens for transformation

Oryza sativa

-

681609

gene PFKFB4, DNA and amino acid sequence determination, expression analysis in DB-1 melanoma cells

Q16877

666334

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

-

664718

KDO-phosphatase knockout, enzyme is encoded by 4 genes, PFKFB1-4, PFKFB3 is activated by mitogenic, inflammatory and hypoxic stimuli

-

673215

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

-

675637

overexpression in Escherichia coli as His6-tagged fusion protein

-

681437

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

-

671260

overexpression of hepatic 6-phosphofructo-2-kinase/fructose-2,6-bisphosphate using an adenoviral vector

-

664763

overexpression of PFK-2/FBPase-2 in MIN6 cells

-

716226

phylogenetic analysis

4 entries

phylogenetic analysis, transcriptional control of isozymes, overview; phylogenetic analysis, transcriptional control of isozymes, overview