Information on EC 2.7.1.147 - ADP-specific glucokinase

Word Map on EC 2.7.1.147
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
2.7.1.147
-
RECOMMENDED NAME
GeneOntology No.
ADP-specific glucokinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ADP + D-glucose = AMP + 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
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
chitin degradation I (archaea)
-
-
Glycolysis / Gluconeogenesis
-
-
glycolysis V (Pyrococcus)
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
glycolysis
-
-
SYSTEMATIC NAME
IUBMB Comments
ADP:D-glucose 6-phosphotransferase
Requires Mg2+. The enzyme from Pyrococcus furiosus is highly specific for D-glucose; there is some activity with 2-deoxy-D-glucose, but no activity with D-fructose, D-mannose or D-galactose as the substrate. No activity is detected when ADP is replaced by ATP, GDP, phosphoenolpyruvate, diphosphate or polyphosphate.
CAS REGISTRY NUMBER
COMMENTARY hide
173585-07-4
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
human ADPGK catalyses ADP-dependent phosphorylation of glucose in vitro
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
ADP + D-fructose
AMP + D-fructose 6-phosphate
show the reaction diagram
ADP + D-fructose 6-phosphate
AMP + D-fructose 1,6-bisphosphate
show the reaction diagram
the enzyme phosphorylates both D-glucose and D-fructose 6-phosphate. Binding of both substrates to the same active site. At a sugar concentration of 10 mM the activity with D-fructose 6-phosphate is about 75% compared to the activity with D-glucose. No activity in presence of ATP. kcat/KM for the phosphorylation of D-fructose 6-phosphate is 440fold higher than the kcat/Km for the phosphorylation of glucose
-
-
ir
ADP + D-glucosamine
AMP + D-glucosamine 6-phosphate
show the reaction diagram
ADP + D-glucose
AMP + D-glucose 6-phosphate
show the reaction diagram
ADP + D-mannose
AMP + D-mannose 6-phosphate
show the reaction diagram
AMP + D-glucose 6-phosphate
ADP + D-glucose
show the reaction diagram
-
-
-
r
CDP + D-glucose
CMP + D-glucose 6-phosphate
show the reaction diagram
12% of the activity with ADP
-
-
?
D-1,5-anhydroglucitol + ADP
?
show the reaction diagram
D-glucose + ADP
D-glucose 6-phosphate + AMP
show the reaction diagram
GDP + D-glucose
GMP + D-glucose 6-phosphate
show the reaction diagram
TDP + D-glucose
TMP + D-glucose 6-phosphate
show the reaction diagram
about 10% compared to the activity with ADP
-
-
r
UDP + D-glucose
UMP + 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
ADP + D-glucose
AMP + D-glucose 6-phosphate
show the reaction diagram
D-glucose + ADP
D-glucose 6-phosphate + AMP
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
required at 0.5 M for full enzyme activity
Pb2+
-
67% of activity
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
D-fructose 6-phosphate
competitive inhibition of D-fructose 6-phosphate when glucose is used as the variable substrate; strong inhibition of glucokinase activity, competitive inhibion versus D-glucose as variable substrate
D-glucose
EDTA
50 mM, complete inhibition; complete loss of activity; completely abolishes activity
Mg2+
excess Mg2+ is inhibitory, 55% inhibition at 55 mM, 41% at 15 mM, and 22% at 10 mM
NaCl
84% of maximal activity at 0.5 M, 48% of maximal activity at 1 M
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
D-glucosamine
10 mM, 40% activation of glucokinase activity
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008 - 16
ADP
0.065
D-fructose 6-phosphate
pH 6.5, 30C
0.096 - 109
D-glucose
0.1 - 2
D-glucose 6-phosphate
2.9 - 20
GDP
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.56 - 116
ADP
16.5
D-fructose 6-phosphate
Methanococcus maripaludis
Q6LXQ3
pH 6.5, 30C
23
D-glucose
Methanococcus maripaludis
Q6LXQ3
pH 6.5, 30C; pH 7.0, 30C; pH 7.8, 30C
0.45 - 0.68
D-glucose 6-phosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.95 - 8400
ADP
13
253
D-fructose 6-phosphate
Methanococcus maripaludis
Q6LXQ3
pH 6.5, 30C
87
0.005 - 207
D-glucose
35
1.09 - 1.1
D-glucose 6-phosphate
105
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.3 - 9.1
ADP
0.06 - 0.5
AMP
0.226
D-fructose 6-phosphate
pH 7.5, 30C, glucokinase reaction; pH 7.8, 30C
2.9 - 895
D-glucose
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.08
ADP
Methanococcus maripaludis
Q6LXQ3
pH 7.5, 30C, reverse glucokinase reaction
40
D-fructose
Methanococcus maripaludis
Q6LXQ3
pH 7.5, 30Cphosphopfructokinase reaction
0.22
D-fructose 6-phosphate
Methanococcus maripaludis
Q6LXQ3
pH 7.5, 30C, glucokinase reaction
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.007
-
substrate: D-glucose, 50C, pH not specified in the publication, enzyme from lactate-grown cells
0.1
-
substrate: D-glucose, 50C, pH not specified in the publication, enzyme from starch-grown cells
11.7
purified recombinant enzyme, pH 7.0, 37C, substrate D-glucose
17
purified recombinant enzyme, pH 6.0, 37C, substrate D-glucose
29.3
purified recombinant enzyme, pH 9.0, 37C, substrate D-glucose
58.9
-
purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 6.5
bimodal optima at pH 5.8-6.5 and at 8.8-9.0
6
-
assay at
8.8 - 9
bimodal optima at pH 5.8-6.5 and at 8.8-9.0
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
pH 5.0: about 50% of maximal phosphofructokinase activity, pH 7.0: about 65% of maximal phosphofructokinase activity
5.5 - 7.5
pH 5.5: about 50% of maximal phosphofructokinase activity, pH 7.5: about 75% of maximal phosphofructokinase activity
5.5 - 8
pH 5.5: about 50% of maximal activity, pH 8.0: about 30% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
specific activity is 14fold higher in starch-grown cells than in lactate-grown cells
Manually annotated by BRENDA team
-
specific activity is 14fold higher in starch-grown cells than in lactate-grown cells
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Thermococcus litoralis (strain ATCC 51850 / DSM 5473 / JCM 8560 / NS-C)
Thermococcus litoralis (strain ATCC 51850 / DSM 5473 / JCM 8560 / NS-C)
Thermococcus litoralis (strain ATCC 51850 / DSM 5473 / JCM 8560 / NS-C)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
-
gel filtration
51500
1 * 51500, recombinant enzyme, SDS-PAGE
52000
-
1 * 52000
53000
-
1 * 53000, SDS-PAGE
54000
-
gel filtration
55000
-
high pressure liquid chromatography
60300
recombinant enzyme, gel filtration
89000
-
gel filtration
93000
-
gel filtration
100000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
structures reveal a ribokinase-like tertiary fold similar to archaeal orthologues but with significant differences in some secondary structural elements. Both the unliganded and the AMP-bound ADPGK structures are in the open conformation and reveal the presence of a disulfide bond between conserved cysteines that is positioned at the nucleotide-binding loop. The AMP-bound structure defines the nucleotide-binding site with one of the disulfide bond cysteines coordinating the AMP with its main chain atoms
molecular modeling of structure. for binding of ADP, residues M347, I431 and L441 create a hydrophobic pocket around the adenine group. R194 makes a hydrogen bond with alpha and beta phosphates, carbonyl and NH groups from V432 peptide bond make a hydrogen bond with the NH2 group of C6 and the N1 atom of adenine
enzyme in complex with AMP, sitting drop vapour diffusion method, 10 mg/ml protein in 6 mM ADP-beta-S, 30 mM glucose mixed in equal volumes with reservoir solution containing 1.6 M citrate, pH 6.5, 10 mM DTT, equilibration against 0.075 ml of reservoir solution at 25C, a few weeks to 1 month, X-ray diffraction structure determination and analysis at 1.9 A resolution, molecular replacement, modeling
-
purified recombinant N-terminally His-tagged apoenzyme, hanging drop vapour diffusion method, 20C, 10 mg/ml protein in solution is mixed with an equal volume of 0.003 ml of mother liquor containing 9-13% PEG 6000, 0.2 M Li2SO4, and 0.1 M citrate buffer, pH 3.6, heavy atom derivatization with HgCl2, X-ray diffraction structure determination and analysis at 2.0 A resolution, single isomorphous replacement with an anomalous scattering
-
2.3 A resolution, R-factor of 20.4%
-
crystal structures of apo form and holo form, in the presence of D-glucose and the nonhydrolyzable ADP analog adenosine 5'-(3-thio)diphosphate. The conformational changes upon sequential substrate binding can be explained by an almost pure rotation (or a rotation plus a translation) facilitated by residues in the flexible inter-domain connection; sitting-drop vapor-diffusion crystallization
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
-
stable up to
additional information
-
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme is not sensitive to oxygen
-
660831
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
600fold to homogeneity by ultracentrifugation, ion exchange chromatography, ammonium sulfate fractionation, hydrophobic interaction chromatography, gel filtration, hydroxyapatite chromatography, and 2 additional ion exchange chromatographic steps
-
recombinant enzyme from Escherichia coli strain BL21(DE3) by heat treatment at 90C, ammonium sulfate fractionation, ion exchange chromatography, and gel filtration
-
recombinant enzyme from Escherichia coli strain DH5alpha by nickel affinity and ion exchange chromatography, to homogeneity
recombinant N-terminally His-tagged enzyme from Escherichia coli by heat treatment and nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, expression in Escherichia coli strain DH5alpha
expressed in Escherichia coli
expression in Escherichia coli
expression in Escherichia coli strain BL21(DE3)
-
gene PH0589, DNA and amino acid sequence determination and analysis, expression of the N-terminally His-tagged in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D481A
greatly reduced activity
D481E
greatly reduced activity
D481N
greatly reduced activity
D84A
greatly reduced activity
H264A
greatly reduced activity, no substrate inhibition with glucose concentrations of up to 55 mM
H382A
no catalytic activity
H382V/H387V
no catalytic activity
H387A
no catalytic activity
R228A
greatly reduced activity
C174S
-
site-directed mutagenesis, activity and oligomarization state are similar to the wild-type enzyme
C94S
-
site-directed mutagenesis, activity is similar to the wild-type enzyme, the mutant enzyme forms no dimers, but monomers
C94S/C174S
-
site-directed mutagenesis, activity is similar to the wild-type enzyme, the mutant enzyme forms no dimers, but monomers
D451A
-
site-directed mutagenesis, nearly inactive mutant
D451N
-
site-directed mutagenesis, nearly inactive mutant
D451S
-
site-directed mutagenesis, nearly inactive mutant
E279D
-
mutation does not affect the KM value for MgADP-, but causes a large increase on KM value for glucose and an 87fold weaker binding of glucose onto the non-hydrolysable enzyme-AMP-AlF3 complex. Mutant is still inhibited by free Mg2+
E279L
-
mutation does not affect the KM value for MgADP-, but causes a large increase on KM value for glucose and an 87fold weaker binding of glucose onto the non-hydrolysable enzyme-AMP-AlF3 complex. Mutant is still inhibited by free Mg2+
E279Q
-
mutation does not affect the KM value for MgADP-, but causes a large increase on KM value for glucose and an 87fold weaker binding of glucose onto the non-hydrolysable enzyme-AMP-AlF3 complex. Mutant is still inhibited by free Mg2+
E308Q
-
mutation increases the KM value for MgADP-, mutant is activated by free Mg2+
E279D
-
mutation does not affect the KM value for MgADP-, but causes a large increase on KM value for glucose and an 87fold weaker binding of glucose onto the non-hydrolysable enzyme-AMP-AlF3 complex. Mutant is still inhibited by free Mg2+
-
E279L
-
mutation does not affect the KM value for MgADP-, but causes a large increase on KM value for glucose and an 87fold weaker binding of glucose onto the non-hydrolysable enzyme-AMP-AlF3 complex. Mutant is still inhibited by free Mg2+
-
E279Q
-
mutation does not affect the KM value for MgADP-, but causes a large increase on KM value for glucose and an 87fold weaker binding of glucose onto the non-hydrolysable enzyme-AMP-AlF3 complex. Mutant is still inhibited by free Mg2+
-
E308Q
-
mutation increases the KM value for MgADP-, mutant is activated by free Mg2+
-