Literature summary for 3.1.3.46 extracted from

Kim, S.G.; Manes, N.P.; El-Maghrabi, M.R.; Lee, Y.H.
Crystal structure of the hypoxia-inducible form of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB3): a possible new target for cancer therapy (2006), J. Biol. Chem., 281, 2939-2944.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged inducible isozyme PFKFB3 in Escherichia coli strain BL21(DE3)	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
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	Homo sapiens
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	Homo sapiens

Crystallization (Comment)

Organism

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

Homo sapiens

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

Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
beta-D-fructose 2,6-bisphosphate + H2O	Homo sapiens	regulation mechanism, overview	D-fructose 6-phosphate + phosphate	-	?
additional information	Homo sapiens	inducible isozyme PFKFB3 plays a crucial role in the progression of cancerous cells by enabling their glycolytic pathways, overview	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P16118	recombinant	-
Homo sapiens	P16118	isozymes PFKFB1-4	-

Purification (Commentary)

Purification (Comment)	Organism
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	Homo sapiens

Reaction

Reaction	Comment	Organism	Reaction ID
beta-D-fructose 2,6-bisphosphate + H2O = D-fructose 6-phosphate + phosphate	substrate binding loop structure and regulation mechanism, interactions of enzyme and substrate involve residues K47, R74, R75, R98, and T126, overview	Homo sapiens	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
liver	isozyme PFKFB1	Homo sapiens	-
testis	isozyme PFKFB4	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
beta-D-fructose 2,6-bisphosphate + H2O	regulation mechanism, overview	Homo sapiens	D-fructose 6-phosphate + phosphate	-	?
beta-D-fructose 2,6-bisphosphate + H2O	substrate binding loop structure, overview	Homo sapiens	D-fructose 6-phosphate + phosphate	-	?
additional information	inducible isozyme PFKFB3 plays a crucial role in the progression of cancerous cells by enabling their glycolytic pathways, overview	Homo sapiens	?	-	?

Synonyms

Synonyms	Comment	Organism
6-phosphofructo-2-kinase/fructose-2,6-bisphosphase	-	Homo sapiens
6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase	-	Homo sapiens
More	bifunctional enzyme, cf. EC 2.7.1.105	Homo sapiens
PFKFB	-	Homo sapiens
PFKFB3	-	Homo sapiens