Information on EC 2.7.4.8 - guanylate kinase

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

EC NUMBER
COMMENTARY hide
2.7.4.8
-
RECOMMENDED NAME
GeneOntology No.
guanylate kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + GMP = ADP + GDP
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
Phosphorylation
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
guanosine ribonucleotides de novo biosynthesis
-
-
Metabolic pathways
-
-
purine metabolism
-
-
Purine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:(d)GMP phosphotransferase
dGMP can also act as acceptor, and dATP can act as donor.
CAS REGISTRY NUMBER
COMMENTARY hide
9026-59-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain JE24F+, derived from W3110, uninfected or infected with RNA-phage MS2
-
-
Manually annotated by BRENDA team
strain K12
Uniprot
Manually annotated by BRENDA team
strain TS202A, guanylate-kinase-deficient
-
-
Manually annotated by BRENDA team
Jerusalem artichoke
-
-
Manually annotated by BRENDA team
strain H37Rv, Rv1389c gene
UniProt
Manually annotated by BRENDA team
nuclear gene VIRESCENT 2 or v2
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
aciclovir (ACV-MP)
?
show the reaction diagram
-
-
-
-
?
ATP + (R)-3-((2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)methoxy)-4-hydroxybutylphosphonic acid
(R)-ganciclovir phosphonate monophosphate
show the reaction diagram
ATP + 6-thioguanosine 5'-monophosphate
ADP + 8-thioguanosine 5'-diphosphate
show the reaction diagram
ATP + 8-azaguanosine 5'-monophosphate
ADP + 8-azaguanosine 5'-diphosphate
show the reaction diagram
ATP + 8-bromoguanosine 5'-monophosphate
ADP + 8-bromoguanosine 5'-diphosphate
show the reaction diagram
-
poor substrate
-
-
?
ATP + 9-(1,3-dihydroxy-2-propoxymethyl)guanine 5'-monophosphate
ADP + 9-(1,3-dihydroxy-2-propoxymethyl)guanine 5'-diphosphate
show the reaction diagram
-
no substrate: 9-(5,5-difluoro-5-phosphonopentyl)guanine 5'-monophosphate
-
-
?
ATP + 9-(2-hydroxyethoxymethyl)guanine 5'-monophosphate
ADP + 9-(2-hydroxyethoxymethyl)guanine 5'-diphosphate
show the reaction diagram
-
i.e. acyclovir 5'-monophosphate
-
-
?
ATP + 9-(5-phosphonopenthyl)guanine
?
show the reaction diagram
-
9-(5,5'-difluoro-5-phosphonopenthyl)guanine is not a substrate
-
-
?
ATP + AMP
ADP + ADP
show the reaction diagram
-
very low activity with AMP
-
-
?
ATP + dGMP
ADP + dGDP
show the reaction diagram
ATP + ganciclovir
ADP + ganciclovir phosphate
show the reaction diagram
-
-
-
-
?
ATP + ganciclovir monophosphate
?
show the reaction diagram
-
-
-
-
?
ATP + GDP
ADP + GMP
show the reaction diagram
-
-
?
ATP + GDP
ADP + GTP
show the reaction diagram
ATP + GDP
ADP + guanosine 5'-tetraphosphate
show the reaction diagram
-
-
?
ATP + GMP
ADP + GDP
show the reaction diagram
ATP + IMP
ADP + IDP
show the reaction diagram
-
very poor substrate
-
-
?
dATP + dGMP
dADP + dGDP
show the reaction diagram
dATP + GMP
dADP + GDP
show the reaction diagram
dGMP + ATP
dGDP + ADP
show the reaction diagram
GMP + ATP
GDP + ADP
show the reaction diagram
GMP + MgATP2-
MgADP- + GDP
show the reaction diagram
guanosine monophosphate + adenosine triphosphate
guanosine diphosphate + adenosine diphosphate
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
aciclovir (ACV-MP)
?
show the reaction diagram
-
-
-
-
?
ATP + dGMP
ADP + dGDP
show the reaction diagram
ATP + ganciclovir
ADP + ganciclovir phosphate
show the reaction diagram
-
-
-
-
?
ATP + GMP
ADP + GDP
show the reaction diagram
dGMP + ATP
dGDP + ADP
show the reaction diagram
GMP + ATP
GDP + ADP
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Li+
-
partial activity
Ni2+
-
activation
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5,5'-dithiobis(2-nitrobenzoate)
6-Selenoguanosine 5'-phosphate
-
GMP-phosphorylation, pI 4.9-isozyme, competitive with respect to GMP
6-Thioguanosine 5'-phosphate
8-azaguanosine 5'-monophosphate
-
(d)GMP-phosphorylation
8-bromoguanosine 5'-monophosphate
-
(d)GMP-phosphorylation
9-(1,3-dihydroxy-2-propylmethyl)guanine 5'-monophosphate
-
-
9-(2-hydroxyethoxymethyl)guanine 5'-monophosphate
-
-
9-(6,6-difluoro-6-phosphonohexyl)guanine
-
competitive with respect to GMP, non-competitive with respect to ATP
-
9-(6-phosphonohexyl)guanine
-
competitive with respect to GMP, non-competitive with respect to ATP
-
AMP
-
10-15% inhibition at 5 mM
Ap5G
Ap5G locks an incompletely closed conformation of the enzyme, in which the adenine moiety is located outside its expected binding site. Instead, it binds at a subunit interface that is unique to the bacterial enzyme, which is in equilibrium between a dimeric and an hexameric form in solution.
Ca2+
-
in the presence of Mg2+
CMP
-
10-15% inhibition at 5 mM
Cs+
-
strong
dAMP
-
10-15% inhibition at 5 mM
dCMP
-
10-15% inhibition at 5 mM
GTP
-
GMP-phosphorylation
indol-3-acetic acid
-
GMP + ATP protect
N-ethylmaleimide
p-chloromercuribenzoic acid
p-hydroxymercuribenzoate
phosphate
-
weak
UMP
-
10-15% inhibition at 5 mM
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.052
(R)-ganciclovir phosphonate
-
pH 7.5, 37C
2.1
6-thioguanosine 5'-monophosphate
-
pH 7.5, 30C
0.013 - 0.091
8-azaguanosine 5'-monophosphate
0.25
9-(5-phosphonopentyl)guanine
-
pH 7.5, 30C
23.6
Adenosine triphosphate
-
0.12 - 1
ATP
1.25
Co2+
-
-
0.01 - 0.4
dGMP
0.047
ganciclovir monophosphate
-
pH 7.5, 37C
0.045 - 0.054
ganciclovir-MP
-
-
0.097
GDP
-
pH 7.5, 25C
0.002 - 1.8
GMP
0.0051
guanosine monophosphate
-
10.8 - 347
K+
-
-
0.017
MgADP2-
-
pH 7.5, 25C
-
0.2 - 0.45
MgATP2-
0.75
Mn2+
-
-
39
NH4+
-
-
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
90
ATP
Saccharomyces cerevisiae
-
pH 7.7, 25C, reverse reaction
2 - 51
dGMP
394 - 1121
GMP
130
GTP
Bos taurus
-
calculated as GDP produced
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.043 - 0.067
9-(1,3-dihydroxy-2-propylmethyl)guanine 5'-monophosphate
0.45 - 0.84
9-(2-hydroxyethoxymethyl)guanine 5'-monophosphate
0.14
9-(6,6-difluoro-6-phosphonohexyl)guanine
-
pH 7.5, 30C, higher affinity than 9-(6-phosphonohexyl)guanine for guanylate kinases al pH values below 7.5
-
0.11
9-(6-phosphonohexyl)guanine
-
pH 7.5, 30C
-
0.148 - 0.57
dGMP
0.215
GDP
-
pH 7.5, 25C
0.035 - 0.63
GMP
0.037
MgADP-
-
pH 7.5, 25C
0.08 - 0.16
MgATP2-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0005
Ap5G
Escherichia coli
P60546
The crystal structure of EcGMPK in complex with Ap5G solved at 2.5 resolution.
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0012
-
E72Q mutant
0.0032
-
D103N mutant, adenylate kinase activity
0.0057
-
E72Q mutant, adenylate kinase activity
0.024
-
E72Q/D103N mutant
0.088
-
D103N mutant
0.44
-
-
1.24
-
-
2.5
-
mutant S35P; mutant S35P/D101S
30
-
with substrates ATP and dGMP
323
-
rod outer segment
354
-
retina
750
-
with substrate ATP and GMP
999
290 Vmax for adenosine triphosphate (ATP); 92 Vmax for guanosine monophosphate (GMP)
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 10
-
dGMP-phosphorylation
7 - 10.9
-
GMP-phosphorylation
7 - 9
-
equal activity in Tris-HCl buffer and 3,3-dimethylglutarate buffer
7.3 - 8.2
-
Tris-chloride buffer
8
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10.9
-
about half-maximal activity at pH 5 and maximal activity at pH 10.9, GMP-phosphorylation, calf thymus
5.2 - 7.5
-
about half-maximal activity at pH 5.2 and 7.5
5.7 - 10.9
-
about half-maximal activity at pH 5.7 and 10.9, dGMP-phosphorylation, calf thymus
6.5 - 9
-
70% of maximal activity at pH 6.5 and 9
6.8 - 8.6
-
about 80% of maximal activity at pH 6.8 and 8.6, about 95% of maximal activity at pH 7.3 and 8
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.7 - 4.8
4.9
-
4 isoelectric variants, agarose gel electrophoresis and isoelectric focusing
5.8
-
pH 7.7, 25C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
strain L60TM, a subline of Earle's L-strain, i.e. L-cells
Manually annotated by BRENDA team
-
Morris 7793 or Dunning
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
outer segment
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
plastid/mitochondrial isozyme
Manually annotated by BRENDA team
-
cytosolic isozyme
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Anaplasma phagocytophilum (strain HZ)
Coxiella burnetii (strain RSA 493 / Nine Mile phase I)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Listeria monocytogenes serovar 1/2a (strain ATCC BAA-679 / EGD-e)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Plasmodium vivax (strain Salvador I)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Staphylococcus aureus (strain COL)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20500
-
gel filtration
21700
-
deduced from the amino acid composition
21900
deduced from the amino acid composition
22000
-
calculated
22010
deduced from the amino acid composition and detected by mass spectrometry, adducts with sulfate and not phosphate are detected in mass spectrometry
23460
monomer, electrospray ionization-mass spectrometry
24000
-
guanylate monophosphate kinase
25000
-
gel filtration
72000
-
gel filtration
88000
-
equilibrium centrifugation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 22000, SDS-PAGE
hexamer
monomer
monomer or dimer
the recombinant and refolded CaVbeta1b-GK is a mixture of dimers and monomers
oligomer
x * 23462, thermodynamic analysis, the enzyme is in equilibrium between a dimer and higher order oligomers, whose relative amounts depend on protein concentration, ionic strength, and the presence of ATP
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
vapour diffusion method
SAP97 SH3-guanylate kinase/p-LGN18 complex, hanging drop vapor diffusion method, using 0.2M ammonium nitrate, 20% (w/v) polyethylene glycol 3350
-
crystal structure of a complex with ADP and GMP
-
SAP97 SH3-guanylate kinase/p-LGN18 complex, hanging drop vapor diffusion method, using 0.2M ammonium nitrate, 20% (w/v) polyethylene glycol 3350
crystal structure of a complex with GMP
-
crystal structure of the enzyme with a non-acetylated N terminus in its unligated form as well as in complex with GMP
octahedral bipyramids, preliminary X-ray analysis
-
temperature-dependent space-group transitions between orthorhombic and tetragonal forms
-
Crystals grown in 20% PEG 3350, 0.2 M LiSO4 and 0.1 M Tris-HCl pH 8.
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 9
-
4C, inactivation within 48 h outside this range
642690
5.5 - 8.5
-
at least 15 min stable at 30C
642688
7.5
-
most stable at
642690
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
at least 15 min stable from pH 5.5-8.5
45
-
t1/2: 10 min
50
-
10 min, inactivation
60
-
10 min, 90% loss of activity
80
-
incubation for 2 min, 5 min, 10 min or 20 min leads to 43%, 67%, 89% or 97% loss of activity, respectively, 30 min: inactivation
100
-
10 min, about 80% loss of activity, t1/2: 3 min (dGMP-phosphorylation), t1/2: 4 min (GMP-phosphorylation)
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol does not stabilize
-
20% glycerol stabilizes: 90% original activity retained
-
bovine serum albumin stabilizes during purification
-
dialysis against water, pH 6 with several changes of outer fluid inactivates, stable to dialysis for less than 36 h without changing the outer fluid
-
dilution inactivates calf thymus enzyme, KCl protects, not bovine serum albumin
-
freeze-thawing rapidly inactivates
-
KCl does not prevent heat inactivation
-
unstable in dilute solutions, below 0.005 mg/ml, bovine serum albumin or other suitable proteins protect, highly purified enzyme is stabilized by the presence of lactic dehydrogenase and pyruvate kinase
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10C, more than a month
-
-15C, more than 6 months
-
-20C, stable at all stages of purification
-
-20C, stable for several months after purification by Blue Sepharose and gel filtration
-25C, stable
-
-30C, in N-ethylmorpholine-HCl buffer, pH 7.5, several years
-
0-4C, 6-10 weeks
-
0C, below, in 70% saturated ammonium sulfate, several years
4C, at pH-values below 4 or above 9, inactivation within 48 h
-
4C, calf thymus enzyme, stable in the absence of thiols
-
4C, diluted calf thymus enzyme solution, 0.002 mg/ml, inactivation within 10 days, KCl protects
-
4C, in 20% glycerol, at least 1 month
-
4C, in N-ethylmorpholine-HCl buffer, pH 7.5, several years
-
4C, partially purified calf thymus enzyme preparation in the presence of 1 M KCl, 3 months
-
glycerol, 20%, stabilizes labile enzyme during storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
4 isoenzymes purified by a method that includes GMP agarose chromatography and isoelectric focusing
-
method
method that includes DEAE-cellulose and Sephadex-75 chromatography
method that includes DEAE-cellulose, hydroxyapatite and Sephadex G-100 chromatography
-
method that includes DEAE-cellulose, hydroxyapatite and Sephadex-75 chromatography
-
method that includes DEAE-cellulose, Sephadex-75 chromatography and isoelectric focusing
-
method that includes DEAE-Sephacel and Blue Sepharose CL 6B chromatography
-
method that includes DEAE-Sephacel, Cibacron-blue Sepharose and two Sephadex-75 chromatography
-
method that includes Sephadex and two DEAE-cellulose chromatography
-
Ni2+-NTA agarose affinity column chromatography
Ni2+-NTA agarose affinity column chromatography and gel filtration
-
partial
partial by a method that includes DEAE-cellulose chromatography
-
partial by a method that includes DEAE-cellulose, hydroxyapatite and Sephadex G-200 chromatography
-
recombinant ecGMPK is purified by a two-step chromatography procedure involving affinity chromatography on Blue Sepharose and gel filtration
recombinant His6-tagged PSD-95 SH3-GK from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, recombinant GST-tagged PSD-95 SH3-GK from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography
-
recombinant protein
TALON metal affinity resin column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis, sequence comparison
-
expressed in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli JM109 cells
-
expression in Escherichia coli
-
expression in Escherichia coli strain BLI5
expression of fluorescence-tagged S-SCAM in COS-7 cells
-
expression of GFP-tagged enzyme in COS cells, expression of His6-tagged and GST-tagged PSD-95 SH3-GK in Escherichia coli strain BL21(DE3)
-
overexpression of Cavbeta1b in Xenopus laevis oocytes in inclusion bodies; overexpression of Cavbeta2a in Xenopus laevis oocytes in inclusion bodies
PSD-95, containing SH3GK residues 417-724 and CASK, containing SH3GK residues 591-909, expression in Escherichia coli
-
transient expression in HEK293 cells, with N-methyl-D-aspartate receptor, wild-type and mutant, cell surface co-expression
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
there is an approximately 40% drop in guanylate kinase mRNA expression in cell lines resistant to 9-[2-(phosphonomethoxyethyl)guanine] and 9-[2-(phosphonomethoxyethyl)diaminopurine]
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C3S/C5S
-
PSD-95aC3S/C5S mutant
S35N/V168F
-
the mutations significantly suppress enzyme catalytic activity
D103N
-
active
E72Q
-
no guanylate or adenylate kinase activity
E72Q/D103D
-
no active
D74A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E101D
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
N103D
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y76F
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
P564S/S631D
-
PSD-95 construct
S35P
-
reduced activity
S35P/D101S
-
reduced activity
S80A
-
sluggish enzyme
Y78F
-
affinity for MgATP2- similar to wild type but affinity for GMP decreases by a factor of 12
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
complete denaturation of the protein is not reversible due to aggregation at the unfolded state
recombinant Cavbeta1b after overexpression in inclusion bodies in Xenopus laevis oocytes; recombinant Cavbeta2a from inclusion bodies after overexpression in Xenopus laevis oocytes
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
medicine
GMPK from bacterial pathogens, in which this enzyme is essential, are potential targets for therapeutic inhibition.
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