Information on EC 2.7.1.2 - glucokinase

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

EC NUMBER
COMMENTARY hide
2.7.1.2
-
RECOMMENDED NAME
GeneOntology No.
glucokinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + D-glucose = ADP + D-glucose 6-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
1,3-propanediol biosynthesis (engineered)
-
-
Amino sugar and nucleotide sugar metabolism
-
-
Bifidobacterium shunt
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Galactose metabolism
-
-
GDP-glucose biosynthesis
-
-
glucose and glucose-1-phosphate degradation
-
-
glycogen degradation I
-
-
Glycolysis / Gluconeogenesis
-
-
glycolysis III (from glucose)
-
-
heterolactic fermentation
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
Neomycin, kanamycin and gentamicin biosynthesis
-
-
Starch and sucrose metabolism
-
-
Streptomycin biosynthesis
-
-
sucrose biosynthesis II
-
-
sucrose degradation III (sucrose invertase)
-
-
trehalose degradation I (low osmolarity)
-
-
trehalose degradation II (trehalase)
-
-
trehalose degradation IV
-
-
trehalose degradation V
-
-
UDP-N-acetyl-D-galactosamine biosynthesis II
-
-
glycogen metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:D-glucose 6-phosphotransferase
A group of enzymes found in invertebrates and microorganisms that are highly specific for glucose.
CAS REGISTRY NUMBER
COMMENTARY hide
9001-36-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
several strains
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
A2
-
-
Manually annotated by BRENDA team
OMZ70, ATCC 33535
-
-
Manually annotated by BRENDA team
no isozymes
SwissProt
Manually annotated by BRENDA team
strain PCC 6803, gene sll0593
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
6-N-(carboxyethyl)ATP + D-glucose
6-N-(carboxyethyl)ADP + D-glucose 6-phosphate
show the reaction diagram
6-N-(carboxymethyl)ATP + D-glucose
6-N-(carboxymethyl)ADP + D-glucose 6-phosphate
show the reaction diagram
6-N-(succinyl)ATP + D-glucose
6-N-(succinyl)ADP + D-glucose 6-phosphate
show the reaction diagram
6-N-[N-(6-aminohexhyl)carbamoyl]ATP + D-glucose
6-N-[N-(6-aminohexhyl)carbamoyl]ADP + D-glucose 6-phosphate
show the reaction diagram
adenosine 5'-triphosphate-polyamidoamine dendrimer + D-glucose
adenosine 5'-diphosphate-polyamidoamine dendrimer + D-glucose 6-phosphate
show the reaction diagram
-
-
-
-
ir
ADP + D-glucose
AMP + D-glucose 6-phosphate
show the reaction diagram
ATP + 2-deoxy-D-glucose
ADP + 2-deoxy-D-glucose 6-phosphate
show the reaction diagram
ATP + beta-D-glucose
ADP + beta-D-glucose 6-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-fructose
ADP + D-fructose 6-phosphate
show the reaction diagram
ATP + D-glucosamine
ADP + D-glucosamine 6-phosphate
show the reaction diagram
-
no reaction with the recombinant enzyme
-
-
-
ATP + D-glucose
ADP + D-glucose 6-phosphate
show the reaction diagram
ATP + D-mannose
ADP + D-mannose 6-phosphate
show the reaction diagram
ATP + mannoheptulose
?
show the reaction diagram
-
sugar binding to recombinant wild-type and mutant glucokinase monitored by kinetic measurement and tryptophan fluorescence
-
-
?
ATP + N-acetyl-alpha-D-glucosamine
ADP + N-acetyl-alpha-D-glucosamine 6-phosphate
show the reaction diagram
CTP + D-glucose
ADP + D-glucose 6-phosphate
show the reaction diagram
38% of the velocity with ATP
-
-
?
CTP + D-glucose
CDP + D-glucose 6-phosphate
show the reaction diagram
D-glucose + ATP
ADP + D-glucose 6-phosphate
show the reaction diagram
-
-
-
?
GTP + D-glucose
GDP + D-glucose 6-phosphate
show the reaction diagram
ITP + D-glucose
IDP + D-glucose 6-phosphate
show the reaction diagram
UTP + D-glucose
UDP + D-glucose 6-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
ATP + D-glucose
ADP + D-glucose 6-phosphate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CTP
-
5% of the activity with ATP
GTP
-
12% of the activity with ATP
ITP
-
20% of the activity with ATP
UTP
-
12% of the activity with ATP
additional information
-
adenosine 5'-triphosphate-polyamidoamine dendrimer, polymer-bound ATP by direct coupling, kinetics and initial reaction rates differ from ATP
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cd2+
-
can partially replace Mn2+ in activation
Fe2+
-
48% of the activation with Mg2+
KCl
-
at 20 mM
Sr2+
-
11% of the activation with Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
-
-
2-deoxy-2-([[(1,1-dioxido-1-benzothiophen-2-yl)methoxy]carbonyl]amino)-beta-D-glucopyranose
-
-
2-deoxy-2-[[3-(4-hydroxyphenyl)propanoyl]amino]-alpha-D-glucopyranose
-
186fold selectivity for Trypanosoma cruzi over human enzyme
2-[[(benzyloxy)carbonyl]amino]-2-deoxy-beta-D-glucopyranose
-
245fold selectivity for Trypanosoma cruzi over human enzyme
6-Amino-6-deoxy-D-glucose
-
noncompetitive to ATP and competitive to glucose
adenosine 5'-diphosphate-polyamidoamine dendrimer
-
product inhibition
-
beta,gamma-methyleneadenosine 5'-thiophosphate
-
noncompetitive to ATP and uncompetitive to glucose
D-fructose 6-phosphate
-
weak
D-glucose 6-phosphate
GK regulatory protein
-
-
-
glucokinase regulatory protein
-
glucokinase-regulatory protein
-
GK regulatory protein, relative inhibition of glucokinase activity through GKRP alone wild-type: 32.5% and GKRP plus 10 microM sorbitol 6-phosphate: 55
-
human glucokinase regulatory protein
-
inhibition is reversed by activator RO-28-1675
-
N-acetyl-alpha-D-glucosamine
-
-
N-acetylglucosamine
N-ethylmaleimide
-
-
p-chlormercuribenzoate
-
-
p-hydroxymercuribenzoate
-
-
palmitoyl-CoA
-
glucokinase is inhibited by endogenous long-chain fatty acyl-CoA in islets from omega3-depleted rats
Trehalose 6-phosphate
-
XTP
-
in presence of ATP
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2R)-3-cyclopentyl-2-[4-(methylsulfonyl)phenyl]-N-(1,3-thiazol-2-yl)propanamide
-
activator associates with glucokinase in a glucose-independent fashion. Kinetic assays reveal a lag in enzyme progress curves that is systematically reduced when the enzyme is preincubated with the activator. Activator binding is enthalpically driven. The kcat value of glucokinase is almost fully limited by product release, both in the presence and absence of activator
(S)-3-(4-((3-fluoropyrrolidin-1-yl)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide
-
activation: 1.9fold
(S)-3-(4-((3-fluoropyrrolidin-1-yl)sulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 2.2fold
(S)-6-(3-cyclopentyl-2-[4-(trifluoromethyl)-1H-imidazol-1-yl]propanamido)nicotinic acid
-
in the presence of liver-specific GKA, progress curves at 1 mM glucose are similar to those at 5 mM, reflecting activation of GK. With steady-state kinetic methods it is shown that there are at least two kinetically distinct forms of glucokinase that interconvert through a slow conformational change and that this interconversion is affected by glucose concentration and a liver-specific GKA
(Z)-2-(4-(cyclopropylsulfonyl)phenyl)-N-(5-(2-methylpropylidene)-4-oxo-4,5-dihydro thiazol-2-yl)-3-(tetrahydro-2Hpyran-4-yl)propanamide
-
activation: 1.17fold
(Z)-N-(5-benzylidene-4-oxo-4,5-dihydrothiazol-2-yl)-2-(4-(cyclopropylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.24fold
1-[6'-(2-hydroxy-2-methylpropoxy)-4-[(5-methylpyridin-3-yl)oxy]-3,3'-bipyridin-6-yl]-3-methylurea
i.e. AM-2394. Compound activates glucokinase with an EC50 of 60 nM, increases the affinity of glucokinase for glucose by approximately 10fold, exhibits moderate clearance and good oral bioavailability in multiple animal models, and lowers glucose excursion following an oral glucose tolerance test in an ob/ob mouse model of diabetes
2-(4-(cyclopropylsulfonyl)phenyl)-3-(tetrahydro-2Hpyran-4-yl)-N-(4,5,6,7-tetrahydrobenzo[d]thiazol-2-yl)propanamide
-
activation: 2.27fold
2-(4-(cyclopropylsulfonyl)phenyl)-N-(4-(prop-1-en-2-yl)thiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 2.4fold
2-(4-(cyclopropylsulfonyl)phenyl)-N-(4-isopropylthiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 2.97fold
2-(4-(cyclopropylsulfonyl)phenyl)-N-(4-oxo-4,5-dihydrothiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.65fold
2-(4-(cyclopropylsulfonyl)phenyl)-N-(5,6-dihydro-4Hcyclopenta[d]thiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 3.4fold
2-(4-(cyclopropylsulfonyl)phenyl)-N-(5-iodo-4-isopropylthiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.34fold
2-(4-(cyclopropylsulfonyl)phenyl)-N-(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c] pyridin-2-yl)-3-(tetrahydro-2Hpyran-4-yl)propanamide
-
activation: 2.26fold
2-(4-(methylsulfonyl)phenyl)-N-(4-phenylthiazol-2-yl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide
-
activation: 1.3fold
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
2-amino-N-(4-methyl-1,3-thiazol-2-yl)-5-[(4-methyl-4H-1,2,4-triazol-3-yl)sulfanyl]benzamide
-
activator associates with glucokinase in a glucose-independent fashion. Kinetic assays reveal a lag in enzyme progress curves that is systematically reduced when the enzyme is preincubated with the activator. Activator binding is enthalpically driven. The kcat value of glucokinase is almost fully limited by product release, both in the presence and absence of activator
2-[4-(methylsulfonyl)phenyl]-3-(tetrahydro-2H-pyran-4-yl)-N-(1,3-thiazol-2-yl)propanamide
-
activation: 2.45fold
3-((3-methylbut-2-en-1-yl)oxy)-5-(4-(morpholinosulfonyl)phenoxy)-N-(thiazol-2-yl)benzamide
-
activation: 2.1fold
3-(4-(((2S,6R)-2,6-dimethylmorpholino)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide
-
activation: 1.5fold
3-(4-(((2S,6R)-2,6-dimethylmorpholino)sulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 1.7fold
3-(4-((4-methoxypiperidin-1-yl)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide
-
activation: 1.7fold
3-(4-(8-oxa-3-azabicyclo[3.2.1]octan-3-ylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide
-
activation: 1.7fold
3-(4-(8-oxa-3-azabicyclo[3.2.1]octan-3-ylsulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 1.9fold
3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(4-oxo-4,5-dihydrothiazol-2-yl)benzamide
-
activation: 1.5fold
3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(5-methylpyrazin-2-yl)benzamide
-
activation: 1.6fold
3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)-N-(thiazol-2-yl)benzamide
-
activation: 2.4fold
3-(4-(cyclopropylsulfonyl)phenoxy)-N-(1,5-dimethyl-1H-pyr-azol-3-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 1.5fold
3-(4-(cyclopropylsulfonyl)phenoxy)-N-(4-(4-fluorophenyl)thiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 0.9fold
3-(4-(cyclopropylsulfonyl)phenoxy)-N-(5-fluorothiazol-2-yl)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 1.8fold
6-[[3-hydroxy-5-(propan-2-yloxy)benzoyl]amino]pyridine-3-carboxylic acid
-
activator associates with glucokinase in a glucose-independent fashion. Kinetic assays reveal a lag in enzyme progress curves that is systematically reduced when the enzyme is preincubated with the activator. Activator binding is enthalpically driven. The kcat value of glucokinase is almost fully limited by product release, both in the presence and absence of activator
N-(4,5-dimethylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.72fold
N-(4-(4-fluorophenyl)thiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.25fold
N-(4-(4-methoxyphenyl)thiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.22fold
N-(4-(tert-butyl)thiazol-2-yl)-3-(4-(cyclopropylsulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 0.7fold
N-(4-cyclopropylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.87fold
N-(4-ethylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide
-
activation: 2.17fold
N-(4-isobutylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide
-
activation: 2.05fold
N-(4-isopropylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide
-
activation: 2.49fold
N-(4-tert-butylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)-propanamide
-
activation: 1.29fold
N-(5-bromo-4-isopropylthiazol-2-yl)-2-(4-(cyclopropylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.67fold
N-(5-fluorothiazol-2-yl)-3-((3-methylbut-2-en-1-yl)oxy)-5-(4-(morpholinosulfonyl)phenoxy)benzamide
-
activation: 1.8fold
N-(5-fluorothiazol-2-yl)-3-(4-((4-methoxypiperidin-1-yl)sulfonyl)phenoxy)-5-((3-methylbut-2-en-1-yl)oxy)benzamide
-
activation: 1.7fold
N-(5-isopropyl-4-methylthiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.13fold
N-(6-fluorobenzo[d]thiazol-2-yl)-2-(4-(methylsulfonyl)phenyl)-3-(tetrahydro-2H-pyran-4-yl)propanamide
-
activation: 1.3fold
RO-0274375
-
synthetic activator, activation of wild-type enzyme and mutants V62A, V62T, and V62L, no activation of mutants V62Q, V62E, V62F, and V62K
-
RO-028165
-
synthetic activator, activation of wild-type enzyme and mutants V62A, V62T, and V62L, no activation of mutants V62Q, V62E, V62F, and V62K
-
RO-0283946
-
synthetic activator, activation of wild-type enzyme and mutants V62A, V62T, and V62L, no activation of mutants V62Q, V62E, V62F, and V62K
-
RO-28-0450
-
racemic, activates the enzyme
RO-28-1675
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.9
2-deoxy-D-glucose
-
30°C, pH 7.5, S151A
18
2-deoxyglucose
-
30°C, pH 7.5, wild-type
0.5 - 0.67
6-N-(carboxyethyl)ATP
0.55 - 0.65
6-N-(carboxymethyl)ATP
0.38 - 1
6-N-(succinyl)ATP
0.5 - 1.25
6-N-[N-(6-aminohexhyl)carbamoyl]ATP
0.15
alpha-D-glucose
-
30°C, pH 7.6
0.00000023 - 12.6
ATP
0.56
CTP
-
30°C, pH 7
4.7 - 240
D-fructose
0.028 - 45
D-glucose
0.08 - 4.4
D-mannose
0.25
GTP
-
30°C, pH 7
0.36
ITP
-
30°C, pH 7
0.19
MgATP2-
-
30°C, pH 7
0.63
UTP
-
30°C, pH 7
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.61 - 1492
ATP
5.9 - 166
beta-D-glucose
0.007 - 1492
D-glucose
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.01 - 11
D-glucose
35
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0041
2-deoxy-2-([[(1,1-dioxido-1-benzothiophen-2-yl)methoxy]carbonyl]amino)-beta-D-glucopyranose
-
His-tagged recombinant protein, pH 7.2, 22°C
0.0013
2-deoxy-2-[[3-(4-hydroxyphenyl)propanoyl]amino]-alpha-D-glucopyranose
-
His-tagged recombinant protein, pH 7.2, 22°C
0.00071
2-[[(benzyloxy)carbonyl]amino]-2-deoxy-beta-D-glucopyranose
-
His-tagged recombinant protein, pH 7.2, 22°C
0.0000128 - 0.000113
GK regulatory protein
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0001
-
fetal glucokinase
0.0003
-
adult glucokinase
0.0018
-
strain KT2440, grown on citrate
0.0164
-
strain KT440 in presence of glucose and toluene
0.0211
-
strain KT2440, grown on glucose
0.0273
-
strain KT440 in presence of glucose
0.26
-
hepatocytes
0.34
-
S151G, D-mannose as substrate
0.57
-
rat hepatocytes treated with adenovirus containing the entire coding sequence of rat liver glucokinase
0.6
-
S151G, D-glucose as substrate
0.786
substrate: D-fructose
0.941
subatrste: D-mannose
1.09
-
purified enzyme, Vmax
1.4
-
N166R-S151A, D-mannose as substrate
1.67
purified enzyme, Vmax
1.95
substrate: D-glucose
2
-
N166R-S151A, D-glucose as substrate
2.16
-
S151C, D-mannose as substrate
3.18
-
S151G, D-fructose as substrate
3.32
-
S151A, D-mannose as substrate
4.1
-
S151A, 2-deoxy-D-glucose as substrate
6
-
S151C, D-glucose as substrate
6.6
-
S151A, D-glucose as substrate
9.12
-
S151C, D-fructose as substrate
14.5
-
N166R-S151A, D-fructose as substrate
23.8
-
S151A, D-fructose as substrate
47.2
-
recombinant protein
60.8
-
N166R, D-mannose as substrate
68
-
wild-type, D-mannose and 2-deoxy-D-glucose as substrate
80
-
wild-type and N166R, D-glucose as substrate
122
-
wild-type, D-fructose as substrate
166
-
purified recombinant enzyme
310
-
N166R, D-fructose as substrate
370
-
purified recombinant enzyme, Vmax
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
-
assay at
7.4
-
assay at
8.2
-
2 optima: a major peak at pH 7.0 and a minor peak at pH 8.2
8.5 - 9
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
-
pH 6.0: about 55% of activity maximum, pH 9.0: about 75% of activity maximum
6 - 10.5
-
pH 6.0: about 30% of activity maximum, pH 10.5: about 80% of activity maximum
6.8 - 7.7
-
50% activity at pH 6.8 and pH 7.7
additional information
-
development of an assay method stable at pH 6.8-8.5 and 75-90°C, overview
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38 - 98
-
below 10% of maximal activity at 38°C, about 90% of maximal activity at 98°C
additional information
-
development of an assay method stable at pH 6.8-8.5 and 75-90°C, overview
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
glk is a nonessential gene that is constitutively expressed during logarithmic growth and has slightly reduced expression in the stationary phase
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Lactococcus lactis subsp. lactis (strain IL1403)
Lactococcus lactis subsp. lactis (strain IL1403)
Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350)
Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350)
Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350)
Trypanosoma cruzi (strain CL Brener)
Trypanosoma cruzi (strain CL Brener)
Trypanosoma cruzi (strain CL Brener)
Trypanosoma cruzi (strain CL Brener)
Trypanosoma cruzi (strain CL Brener)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24000
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alpha2, 2 * 24000, SDS-PAGE
33000
-
alpha2, 2 * 33000, SDS-PAGE
33500
-
2 * 33500, the recombinant enzyme probably is a dimer, SDS-PAGE
34500
-
2 * 34500, SDS-PAGE
35200
recombinant enzyme, about, mass spectrometry, 2 peaks
36000
-
2 * 36000, alpha2, recombinant enzyme, SDS-PAGE
41000
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gel filtration
43000
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x * 43000, the enzyme may occur as a monomer or dimer, dependent on the protein concentration, SDS-PAGE
47000
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x * 47000, SDS-PAGE
49000
monomer, gel filtration
52000
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x * 52000, SDS-PAGE
65000
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gel filtration
67000
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gel filtration
67500
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density gradient centrifugation
70000
recombinant enzyme, oxidative conditions, PAGE, the wild-type and mutant enzymes occur as both monomer and homodimer
70400
recombinant enzyme, about, mass spectrometry, 2 peaks
75000
-
recombinant enzyme, gel filtration
87000
-
HPLC gel filtration
additional information
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recombinant enzyme, native PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
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glucokinase is modified by O-GlcNAcylation. siRNA-mediated O-linked N-acetylglucosamine transferase knock-down not only decreases O-GlcNAc content but also glucokinase protein level
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant selenomethionine-labeled enzyme free or in complex with D-glucose, hanging drop vapour diffusion method, 0.002 ml enzyme solution containing 6.8 mg/ml protein mixed with 0.004 ml reservoir solution containing 0.1 M Tris-HCl, pH 8.5, 0.2 M MgCl2, and 1.7 M ammonium sulfate for the apo-enzyme or 18.5-20% PEG 6000 with 2 mM D-glucose and 2-3 mM ADP for the glucose-bound enzyme, X-ray diffraction structure determination and analysis at 2.3-2.2 A resolution
homology modeling of structure reveals high structural homology with human glucokinase
first structures of a glucokinase-glucose complex without activator, of glucokinase-glucose-AMP-PNP and of glucokinase-glucose-AMP-PNP with a bound activator are reported. All structures are extremely similar, thus demonstrating that binding of GK activators does not result in conformational changes of the active protein but in stabilization of the active form of glucokinase
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molecular docking of glucose and synthetic ligands (allosteric activators). Predicted position of glucose coincides with its position in the crystallographic complex. Glucose interacts with the residues of large (Ile225, Gly229, Cys230, Asn231, Glu256, Gln287, Glu290) and small (Ser151, Phe152, Pro153, Thr168, Lys169) domains and with the residues of connecting region Asn204, Asp205. Hydrogen bonds with Asp205, Glu256 and Glu290 stabilize this complex. The allosteric site formed by the small and large domains and two loops connecting them is located approximately 20 A away from the glucose binding site. There are two stable positions of glucokinase activators and approximately five sites of ligand binding with less favorable interaction energy. Predicted position of glucose coincides with its position in the crystallographic complex
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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
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homology modeling of structure. Staphylococcus aureus glucokinase exhibits very close homology with Enterococcus faecalis and Clostridium difficle while with other bacteria it show a high degree of variations both in domain and nondomain regions. Glucose interacts with residues V77, N114, S161 and G160 forming a total of eight hydrogen bonds. Glucose docking score is -12.3697 kcal/mol
crystal structures of apo-SgGlkA, SgGlkA in complex with glucose, and SgGlkA in complex with glucose and adenylyl imidodiphosphate (AMPPNP) are reported. SgGlkA is divided into a small alpha/beta domain and a large alpha + beta domain, and it forms a dimer-of-dimer tetrameric configuration
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crystallized using the sitting-drop vapour-diffusion method. A crystal of SgGlkA in complex with glucose is obtained and diffracted X-rays to 1.84 A resolution
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crystal structures in complexes with inhibitors 2-[[(benzyloxy)carbonyl]amino]-2-deoxy-beta-D-glucopyranose, 2-deoxy-2-[[3-(4-hydroxyphenyl)propanoyl]amino]-alpha-D-glucopyranose, 2-deoxy-2-([[(1,1-dioxido-1-benzothiophen-2-yl)methoxy]carbonyl]amino)-beta-D-glucopyranose. The analogues all share a common glucose moietythat preserves key enzyme-substrate hydrogen bonding interactions with the monosaccharide hydroxyl groups from C1, C3, C4,and C6. The C2 hydroxyl is replaced by a NH group and also par-ticipates in the key hydrogen bonding
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crystals of TcGlcK in complex with D-glucose and ADP are obtained by the hanging-drop, vapor-diffusion method, using PEG3350 as precipitant agent and diammonium hydrogen citrate as additive. A complete native dataset is collected to 2.1 A maximum resolution
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
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48 h, with glucose and DTT stable, unstable without
40
-
stabilization by D-glucose and glycerol is limited since thermal glucokinase denaturation above 40 °C is irreversible even in their presence
55
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1 min, complete loss of activity
60
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no loss of activity in 30 min
75
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about 25% loss of activity in 30 min, presence of 0.01 M glucose + 0.2 M NaCl, about 90% loss of activity, presence of 0.01 M glucose absence of NaCl
80
-
quite stable for 120 min
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
D-glucose stabilizes the enzyme through a specific, ligand induced intramolecular transition from an open to a closed conformation
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DTT stabilizes at 70°C
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glucose increases thermal stability
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glucose or DTT protects against thermal inactivation at 25°C
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glycerol increases the stability
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rapid loss of activity in absence of DTT
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Glycerol
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glycerol causes large functional changes and increases stability as a result of generalized restructuring of surface water of glucokinase by an indirect mechanism and these changes can occur with minimal perturbation of the protein folding structure and with preservation of the unique cooperative kinetics of the enzyme whereas D-glucose stabilizes the enzyme through a specific, ligand induced intramolecular transition from an open to a closed conformation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 1 mM DTT, 20-30% glycerol, indefinitely stable
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-20°C, purified recombinant enzyme, 50 mM Tris-HCl, pH 7.5, stable
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-20°C, stable for 4 weeks
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room temperature, 1 mM DTT, 5 mM MgCl2, stable for 1 week , 4°C, suspended precipitate in a 60% saturated solution of ammonium sulfate, stable for at least 3 months
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
glutathione S-transferase (GST)-GlkB fusion proteins, wild-type and mutant
His-tagged recombinant TcGlcK is expressed in the Escherichia coli BL21 strain and purified to homogeneity by nickel affinity chromatography
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recombinant enzyme
recombinant enzyme from Escherichia coli strain BL21(DE3) by heat treatment and Ni2+ chelating affinity chromatography
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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
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recombinant His-tagged enzyme from strain BL21(DE3) by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain RB791 by nickel affinity chromatography
recombinant N-terminally His6-tagged hepatic enzyme from Escherichia coli by nickel affinity chromatography to over 90% purity
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recombinant N-terminally His6-tagged pancreatic enzyme from Escherichia coli by nickel affinity chromatography to over 90% purity
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recombinant soluble His-tagged enzyme from Escherichia coli strain BL21(DE3) by ultracentrifugation, nickel affinity chromatography, and gel filtration to homogeneity
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recombinant wild-type and mutant enzymes from Escherichia coli to homogeneity
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recombinant wild-type and mutant V62M and E300K enzymes as GST-fusion proteins by glutathione affinity chromatography to homogeneity
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using Ni-NTA chromatography
using Ni-NTA chromatography followed by ion-exchange chromatography on a HiTrap Q FF column and gel filtration on a Superdex 200 column
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli as a GST-fusion protein
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expressed in Escherichia coli as a GST-tagged fusion protein
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expressed in Escherichia coli as a His-tagged fusion protein
expression in Escherichia coli
expression in Saccharomyces cerevisiae
expression of genes nanK, yajF, and ycfX in glk-deficient mutant strain BM5340(DE3)
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expression of hepatic wild-type and mutant enzymes in Escherichia coli strain DH5 alpha as FLAG-tagged proteins
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expression of the N-terminally His6-tagged hepatic enzyme in Escherichia coli
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expression of the N-terminally His6-tagged pancreatic enzyme in Escherichia coli
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expression of wild-type and mutant enzymes as GST-fusion proteins
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expression of wild-type and mutant enzymes in Escherichia coli BL21(DE3)
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gene glcK, phylogenetic analysis and tree, functional complementation of enzyme-deficient Escherichia coli strain UE26, overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain RB791 as soluble proteins
gene glk is organized in gene clusters, transcriptomic analysis of KT2440 in response to glucose, overview
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gene glk, expression of His-tagged enzyme in strain BL21(DE3)
gene sll0593, DNA and amino acid sequence determination
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gene TM 1469 or glk, functional expression of the soluble His-tagged in Escherichia coli strain BL21(DE3)
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gene TM 1469, expression in Escherichia coli strain BL21(DE3)
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glcK open reading frame is cloned from Bacillus sphaericus strain C3-41 and then expressed in Escherichia coli
glutathione S-transferase (GST)-GlkB fusion proteins
overexpression in Escherichia coli UE79
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overexpression in rat hepatocytes
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C166A
site-directed mutagenesis, mutant shows activity similar to the wild-type enzyme
C175A
site-directed mutagenesis, inactive active site residue mutant
C177A
site-directed mutagenesis, inactive active site residue mutant
C182A
site-directed mutagenesis, inactive active site residue mutant
C282A
site-directed mutagenesis, mutant shows slightly increased enzyme activity compared to the wild-type enzyme, conformational changes different from the wild-type enzyme, overview
C321A
site-directed mutagenesis, mutant shows 5fold increased enzyme activity compared to the wild-type enzyme, conformational changes different from the wild-type enzyme, overview
D10K
site-directed mutagenesis, ATP binding site mutant, inactive mutant
A379S
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mutation isolated in a type 2 diabetes patient
A379T
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kcat(1/sec): 61.2, D-glucose S05 (mM): 12.3, Km (ATP): 0.87 mM, relative inhibition of glucokinase activity through GKRP alone: 17% and GKRP plus 10 microM sorbitol 6-phosphate: 53%
A379V
mutations is associated with mature-onset diabetes of the young, type 2 (MODY2). Vmax is 65% of maximal activity, Km-value for ATP is 1.5fold higher than wild-type enzyme
C213R
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kcat(1/sec): 50, D-glucose S05 (mM): 21.6, Km (ATP): 0.89 mM, relative inhibition of glucokinase activity through GKRP alone: 19% and GKRP plus 10 microM sorbitol 6-phosphate: 43%
C233R
mutations is associated with mature-onset diabetes of the young, type 2 (MODY2). Mutation affects a critical residue of the active center of the enzyme and rendered a protein with undetectable enzymatic activity
C252Y
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kcat(1/sec): 29.3, D-glucose S05 (mM): 31.6, Km (ATP): 0.75 mM, relative inhibition of glucokinase activity through GKRP alone: 4% and GKRP plus 10 microM sorbitol 6-phosphate: 4%
D158A
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naturally occurring mutation, activity and kinetics are similar to the wild-type enzyme
D400Y
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mutation isolated in a type 2 diabetes patient
D78A/D158A
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site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
E265K
mutations is associated with mature-onset diabetes of the young, type 2 (MODY2). Vmax is 86% of maximal activity, Km-value for ATP is 1.13fold lower than wild-type enzyme
E300A
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mutation isolated in a type 2 diabetes patient
E300K
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site-directed mutagenesis, mutant shows decreased activity and increased thermolability compared to the wild-type enzyme
E339K
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crystal structure of E339K glucokinase in complex with glucose is shown. This mutation results in a conformational change of His416, spatially interfering with adenosine-triphosphate (ATP) binding
E395A
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mutation isolated in a type 2 diabetes patient
E395G
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mutation isolated in a type 2 diabetes patient
E442K
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kcat(1/sec): 52.6, D-glucose S05 (mM): 5.24, Km (ATP): 1.5 mM, relative inhibition of glucokinase activity through GKRP alone: 14% and GKRP plus 10 microM sorbitol 6-phosphate: 40%
G227A/D158A
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site-directed mutagenesis, reduced activity compared to the wild-type enzyme
G68K
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kcat(1/sec): 43.2, D-glucose S05 (mM): 2.34, Km (ATP): 0.39 mM, relative inhibition of glucokinase activity through GKRP alone: 2% and GKRP plus 10 microM sorbitol 6-phosphate: 20.5%
G68V
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kcat(1/sec): 62.7, D-glucose S05 (mM): 2.2, Km (ATP): 0.31 mM, relative inhibition of glucokinase activity through GKRP alone: 15% and GKRP plus 10 microM sorbitol 6-phosphate: 32.5%
G72R
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