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

glucagon

-

94934, 94939, 94941

glycerol 1-phosphate

Glycerol 2-phosphate

GTP

-

ITP

-

MgATP2-

-

MgGTP2-

phosphate

phosphoenolpyruvate

-

Protein kinase C

-

Protein phosphatase 2A

-

cyclic AMP dependent protein kinase inactivation reversible by

94931

-

salicylate

-

additional information

6 entries

-

0.0102 - 0.204

beta-D-fructose 2,6-bisphosphate

0.00013

D-fructose 2,6-bisphosphate

-

pH 7.4

650442

0.000028

D-fructose-1,6-bisphosphate

pH 6.0, 25°C

0.0001 - 1.4

fructose 2,6-bisphosphate

19 entries

additional information

-

0.1

fructose 2,6-bisphosphate

-

94933, 94945

0.08

6-phosphogluconate

-

9.7

D-fructose 6-phosphate

-

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

-

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

1650

phosphatase activity

20 - 40

additional information

5.5

7

7.3

-

7.5

-

assay at

713740

7.6

8

-

assay at

additional information

-

pH-dependence of kinetic parameters

6.5 - 8

-

37

-

assay at

713740

9.29

Desulfovibrio desulfuricans

-

amino acid sequence calculation

-

663935

-

Drosophila melanogaster

6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase long form; gene Pfrx

SwissProt

-

-

-

-

bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase

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C57/BL6

-

-

-

-

-

-

-

Pigeon

-

-

-

-

Rattus norvegicus Wistar

Wistar

678087

-

Schizosaccharomyces pombe

-

94928, 94931

-

-

-

-

SwissProt

-

-

stock 427, isozymes 1-4

-

Trypanosoma cruzi

-

Vigna radiata var. radiata

-

-

-

-

-

-

-

-

-

-

-

-

-

breast carcinoma cell

esophageal squamous cell carcinoma cell

-

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

-

-

-

-

-

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

-

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

-

-

-

-

both PFKFB3 protein and gene expression is significantly higher than in adjacent non-tumor tissues. Both PFKFB3 protein and gene expression are related to infiltration depth, stage, tumor metastasis, and the degree of tumor differentiation in esophageal squamous cell carcinoma

761806

fat pad

non-small cell lung cancer cell

-

epididymal, reduced expression of isozyme PFKFB3

-

-

-

mRNA abundance of PFKFBP1-2 in normoxia is higher in gills than in hepatopancreas but is undetectable in muscle

-

-

-

-

mRNA abundance of PFKFBP1-2 in normoxia is higher in gills than in hepatopancreas but is undetectable in muscle

-

-

-

-

-

-

lung adenocarcinoma cell

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

bloodstream form, grown in rats

-

-

five isoforms of PFKFB4 are expressed in melanoma cells, of which the D and F isoforms are highly constitutive, while the canonical B isoform seems to be the main isoform induced in hypoxia

five isoforms of PFKFB4 are expressed in melanoma cells, of which the D and F isoforms are highly constitutive, while the canonical B isoform seems to be the main isoform induced in hypoxia

772249

additional information

-

-

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

drug target

malfunction

metabolism

9 entries

physiological function

15 entries

additional information

100000

107000

-

equilibrium sedimentation

94935

110000

12000

-

gel filtration

94951

140000

-

isozyme 2, gel filtration

320000

gel filtration

33000

-

2 * 33000, SDS-PAGE

370000

gel filtration

50000

-

2 * 50000, SDS-PAGE

722278

53000

Pigeon

-

2 * 53000, SDS-PAGE

94936

53300

-

53300, SDS-PAGE

94952

53800

Bos taurus

-

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

600000

-

about, isozyme 1, gel filtration

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

?

dimer

homodimer

monomer

-

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

94926

tetramer

additional information

26 entries

phosphoprotein

12 entries

to 1.8 A resolution. Citrate cocrystallizes in the 2-kinase domain, occupying the fructose-6-phosphate binding-site and extending into the gamma-phosphate binding pocket of ATP. In complex with ADP, ADP mimicks the catalytic binding mode of ATP

Bos taurus

P26285

752141

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

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

to 2.0 A resolution. Citrate cocrystallizes in the 2-kinase domain, occupying the fructose-6-phosphate binding-site and extending into the gamma-phosphate binding pocket of ATP. In complex with ATP analogue AMPPNP, the presence of a gamma-phosphate makes adoption of the catalytic ATP binding mode impossible for AMPPNP, forcing the analogue to bind atypically with concomitant conformational changes to the ATP binding-pocket

O60825

752141

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

Sus scrofa

-

680855

H444A/R445A/R447A

-

enhances catalytic rate of bisphosphatase activity and abrogates inhibition by kinase domain

640339

P456E/P457A/A458E

-

change of chicken sequence to rat sequence, enzyme behaves like rat enzyme

650442

A44G

-

site directed mutagenesis

A44V

-

site directed mutagenesis

K168A

-

site directed mutagenesis

K168N

-

site directed mutagenesis

K168R

-

site directed mutagenesis

K472/473A

mutant fails to localize in nucleus, mainly localizes in cytoplasm. The mutant inhibits the autophagic process, stimulates lactate production, and decreases the activity of AMPK compared to the wild-type

751774

E325A

-

role of E325 as acid-base during reaction

649951

E327A

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

11 entries

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 lessonae

-

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

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

-

-

5 entries

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

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)

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

666334

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

-

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

Desulfovibrio desulfuricans

possible evolution of the proteobacterial gene by horizontal gene transfer from an eukaryote, phylogenetic analysis

-

663935

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

-

714090

consistently overexpressed in lung cancer

761491

p53 downregulates PFKFB4 expression by binding to its promoter and mediating transcriptional repression via histone deacetylases

751761

the canonical B isoform seems to be the main isoform induced in hypoxia

772249

the enzyme is induced in hypoxia and upregulated as a consequence of the transcriptional changes orchestrated by the androgen receptor in prostate cancer cells

-

729519

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

730530

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

-

681540

agriculture

Oryza sativa

-

FK2P in rice is encoded by two genes OsF2KP1 and OsF2KP2, both are involved in the regulation of primary carbon metabolism in rice leaves during the light period

681609

diagnostics