Literature summary for 2.7.1.1 extracted from

Kamata, K.; Mitsuya, M.; Nishimura, T.; Eiki, J.; Nagata, Y.
Structural basis for allosteric regulation of the monomeric allosteric enzyme human glucokinase (2004), Structure, 12, 429-438.

Activating Compound

Activating Compound	Comment	Organism	Structure
2-amino-4-fluoro-5-(1-methyl-1H-imidazol-2-ylsulfanyl)-N-thiazol-2-yl-benzamide	potent synthetic allosteric activator, acts on the wild-type and the N-terminal deletion mutants DELTA N1-11 and DELTA N1-15, mechanism of activation	Homo sapiens

Cloned(Commentary)

Cloned (Comment)	Organism
expression of hepatic wild-type and mutant enzymes in Escherichia coli strain DH5 alpha as FLAG-tagged proteins	Homo sapiens

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant hepatic wild-type and deletion mutant enzymes in complex with either D-glucose or the synthetic activator 2-amino-4-fluoro-5-(1-methyl-1H-imidazol-2-ylsulfanyl)-N-thiazol-2-yl-benzamide, hanging drop vapour diffusion method, 10 mg/ml protein in 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 5 mM Tris(2-carboxyethyl)phosphine hydrochloride, and 20 mM D-glucose, or 0.3 mM compound A, 0.0015-0.003 ml of the solution is mixed with an equal volume of precipitant solution containing 28-30% PEG 1500, 0.1 M HEPES-NaOH, pH 6.0, equilibration against 1 ml precipitant solution, 1 week, crystallization of the free hepatic enzyme by using precipitant solution containing 50 mM NaCl, 1.5-1.6 M ammonium sulfate, and 0.1 M bicine-NaOH, pH 8.7, 1 week, X-ray diffraction structure determination and analysis at 2.3 and 3.4 A resolution, respectively, molecular replacement	Homo sapiens

Crystallization (Comment)

Organism

purified recombinant hepatic wild-type and deletion mutant enzymes in complex with either D-glucose or the synthetic activator 2-amino-4-fluoro-5-(1-methyl-1H-imidazol-2-ylsulfanyl)-N-thiazol-2-yl-benzamide, hanging drop vapour diffusion method, 10 mg/ml protein in 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 5 mM Tris(2-carboxyethyl)phosphine hydrochloride, and 20 mM D-glucose, or 0.3 mM compound A, 0.0015-0.003 ml of the solution is mixed with an equal volume of precipitant solution containing 28-30% PEG 1500, 0.1 M HEPES-NaOH, pH 6.0, equilibration against 1 ml precipitant solution, 1 week, crystallization of the free hepatic enzyme by using precipitant solution containing 50 mM NaCl, 1.5-1.6 M ammonium sulfate, and 0.1 M bicine-NaOH, pH 8.7, 1 week, X-ray diffraction structure determination and analysis at 2.3 and 3.4 A resolution, respectively, molecular replacement

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of N-terminal deletion hepatic enzyme mutants lacking 11 or 15 amino acid residues, respectively	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	positive cooperation, mechanism, kinetic model	Homo sapiens

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + D-glucose	Homo sapiens	-	ADP + D-glucose 6-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P35557	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant FLAG-tagged hepatic wild-type and mutant enzymes from Escherichia coli strain DH5 alpha by ion exchange and glucosamine affinity chromatography, and gel filtration	Homo sapiens

Reaction

Reaction	Comment	Organism	Reaction ID
ATP + D-glucose = ADP + D-glucose 6-phosphate	positive cooperation, mechanism	Homo sapiens	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
liver	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + D-glucose	-	Homo sapiens	ADP + D-glucose 6-phosphate	-	?

Subunits

Subunits	Comment	Organism
monomer	crystal structure determination	Homo sapiens
More	determination of active open and inactive closed enzyme conformation and structure, folding patterns, conformational changes, overview	Homo sapiens

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Homo sapiens

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Homo sapiens