Information on EC 4.1.1.32 - phosphoenolpyruvate carboxykinase (GTP)

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

EC NUMBER
COMMENTARY hide
4.1.1.32
-
RECOMMENDED NAME
GeneOntology No.
phosphoenolpyruvate carboxykinase (GTP)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
GTP + oxaloacetate = GDP + phosphoenolpyruvate + CO2
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylation
-
-
-
-
decarboxylation
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
anaerobic energy metabolism (invertebrates, cytosol)
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
Citrate cycle (TCA cycle)
-
-
gluconeogenesis
-
-
gluconeogenesis III
-
-
Glycolysis / Gluconeogenesis
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
Pyruvate metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
GTP:oxaloacetate carboxy-lyase (adding GTP; phosphoenolpyruvate-forming)
ITP can act as phosphate donor.
CAS REGISTRY NUMBER
COMMENTARY hide
9013-08-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
Holstein cows, dairy cows
-
-
Manually annotated by BRENDA team
strain 1224-5/9
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Modiolus demissus
-
-
-
Manually annotated by BRENDA team
strain MC2 155
SwissProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
three groups of animals examined: high energy diet, high energy diet and exercising, and low energy diet, highest activity in the high energy fed group
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain P2
UniProt
Manually annotated by BRENDA team
strain P2
UniProt
Manually annotated by BRENDA team
; strain KOD1, hyperthermophilic archaeon, highest enzyme level when grown with pyruvate as substrate
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
enzyme loss impairs gluconeogenesis from lactate, lowers plasma glucose, insulin, and triglycerides, reduces hepatic glycogen, and increases glycerol turnover
metabolism
-
approximately a third of gluconeogenesis comes from the enzyme
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
2'-deoxyguanosine 5'-triphosphate + oxaloacetate
2'-deoxyguanosine 5'-diphosphate + phosphoenolpyruvate + CO2
show the reaction diagram
-
2-deoxyGTP is a less effective substrate than GTP, 2-deoxyGTP binds to enzyme less tight than 2-deoxyGDP
2-deoxyGDP is a less effective substrate than GDP in the reverse reaction
?
ATP + oxaloacetate
ADP + phosphoenolpyruvate + CO2
show the reaction diagram
dGTP + oxaloacetate
dGDP + phosphoenolpyruvate + CO2
show the reaction diagram
-
-
-
-
?
GDP + (Z)-3-fluorophosphoenolpyruvate + CO2
?
show the reaction diagram
-
-
-
-
-
GDP + phosphoenolpyruvate + CO2
GTP + oxaloacetate
show the reaction diagram
GTP + oxaloacetate
?
show the reaction diagram
GTP + oxaloacetate
GDP + phosphoenolpyruvate + CO2
show the reaction diagram
IDP + phosphoenolpyruvate + CO2
ITP + oxaloacetate
show the reaction diagram
ITP + oxaloacetate
IDP + phosphoenolpyruvate + CO2
show the reaction diagram
oxaloacetate + ?
pyruvate + CO2
show the reaction diagram
-
ir
-
-
UDP + phosphoenolpyruvate + CO2
UTP + oxaloacetate
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
GDP + phosphoenolpyruvate + CO2
GTP + oxaloacetate
show the reaction diagram
GTP + oxaloacetate
?
show the reaction diagram
GTP + oxaloacetate
GDP + phosphoenolpyruvate + CO2
show the reaction diagram
ITP + oxaloacetate
IDP + phosphoenolpyruvate + CO2
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
-
sligthly activates
Ca2+
-
poor activating cation
Co3+
-
study of Co3+-PEPCK with Co3+ bound to enzyme at site n1
Cr3+
-
study of Co3+-PEPCK with Cr3+ bound to enzyme at site n1
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(Z)-Phosphoenol-3-bromopyruvate
-
linear competitive
1-allyl-3-butyl-8-methylxanthine
-
weak, competitive inhibitor with respect to GTP, IC50: 0.225 mM in the decarboxylation direction, modifications at N-1 and C-8 improve the inhibitory activity by more than 100fold
2,3-Butanedione
-
-
2-oxobutanoate
-
weak
2-oxoglutarate
2-Phosphoglycolate
-
-
3-Aminopicolinic acid
Modiolus demissus
-
-
3-Mercaptopicolinic acid
3-Nitro-2-oxopropionic acid
-
weak
5,5'-dithiobis(2-nitrobenzoate)
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
inhibits wild-type enzyme, but not C306A or C306S mutant enzymes
8-Azidoguanosine 5'-triphosphate
-
photoinactivation is caused by formation of an intramolecular cystine disulfide bridge
Acetopyruvate
-
weak noncompetitive
ADP
Modiolus demissus
-
uncompetitive with respect to IDP, partially competitive with respect to phosphoenolpyruvate
alpha-(Dihydroxyphosphinylmethyl) acrylic acid
-
weak linear competitive
alpha-ketoglutarate
inhibits oxaloacetate formation, mixed-type inhibition
beta-sulfopyruvate
citrate
-
above 5.0 mM, weak
Co3+-GDP
-
competitive inhibitor
Co3+-GTP
-
competitive inhibitor
Cr3+-GDP
-
competitive inhibitor
Cr3+-GTP
-
competitive inhibitor
dithiothreitol
-
5 mM, 19% inhibition
DL-3-Nitro-2-hydroxypropionic acid
-
weak
DL-isocitrate
-
above 5.0 mM, weak
EDTA
-
0.5 mM, 86% inhibition
FeCl2
-
1.5 mM FeCl2 causes rapid inactivation. 0.05 mM FeCl2 and 0.0044 mg/ml ferroactivator, enhance to 3.1times the unstimulated rate
glycerate
-
D-glycerate, L-glycerate and DL-glycerate, weak noncompetitive
GTP
-
competitive inhibitor
Hydroxymalonate
Modiolus demissus
-
slight
IMP
-
competitive inhibitor
interleukin-10
-
added to cell culture medium in combination with interleukin-1beta, reduces mRNA and enzyme level
-
Interleukin-1beta
-
added to cell culture medium in combination with interleukin-10, reduces mRNA and enzyme level
-
iodoacetamide
iodoacetate
-
-
KHCO3
-
above 0.1 M inhibits the oxaloacetate-forming activity
malate
malonate
-
weak
Mg2+
-
Mg2+ reduces mitochondrial PEPCK activity
MgCl2
-
1.5 mM, 62% inhibition
oxalate
pyruvate
weak inhibition
quinolinic acid
-
strongly inhibits the enzyme activated by ferroactivator and Fe2+, no inhibition of Mn2+-activated enzyme
succinate
-
above 5.0 mM, weak
Tartronate
-
weak
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
maximum stimulation at 75 mM
dithiothreitol
maximum at 10-40 mM, 30% stimulation
Ferroactivator
-
ferroactivator + Fe2+, stimulates both directions of the reaction
-
ITP
-
stimulates Mn2+-dependent CO2-oxaloacetate exchange
Mn2+
-
activity regulator, 0.7 mM leads to full enzyme stimulation
reduced glutathione
maximum stimulation at 20 mM
thiol-containing reducing agent
activates
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03
(Z)-3-fluorophosphoenolpyruvate
-
-
0.053 - 0.19
2'-deoxyguanosine 5'-diphosphate
0.71 - 1.05
2'-deoxyguanosine 5'-triphosphate
2.23
ADP
pH 7.0, 60C
0.465
ATP
pH 7.0, 60C
0.814 - 8.3
CO2
0.003 - 20.6
GDP
0.0074 - 1.6
GTP
0.0133 - 46
HCO3-
0.013 - 2.5
IDP
0.05 - 4
ITP
4.5
KHCO3
-
enzyme from autotrophically grown cells
0.042
MnGDP2-
-
-
-
0.197
MnIDP-
-
-
-
40
MnITP2-
-
-
0.004 - 2.5
oxaloacetate
0.0185 - 9.6
phosphoenolpyruvate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4 - 4.6
2'-deoxyguanosine 5'-diphosphate
2.3 - 3.4
2'-deoxyguanosine 5'-triphosphate
1 - 19
CO2
3.5 - 19
GDP
10.3 - 54
GTP
2.3 - 6.9
IDP
10.7 - 15.6
ITP
13.3 - 54
oxaloacetate
1 - 19
phosphoenolpyruvate
additional information
additional information
Corynebacterium glutamicum
-
kcat for NaHCO3: 6.7 s-1
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.2 - 16
CO2
14 - 510
GDP
270 - 790
GTP
19 - 1000
oxaloacetate
15 - 66
phosphoenolpyruvate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1864
GMP
-
in Tris-HCl (50 mM, pH 7.4), temperature not specified in the publication
0.07642
GTP
-
in Tris-HCl (50 mM, pH 7.4), temperature not specified in the publication
0.3483
IMP
-
in Tris-HCl (50 mM, pH 7.4), temperature not specified in the publication
0.2264
ITP
-
in Tris-HCl (50 mM, pH 7.4), temperature not specified in the publication
120
KCl
Modiolus demissus
-
-
115
NaCl
Modiolus demissus
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.225
1-allyl-3-butyl-8-methylxanthine
Homo sapiens
-
weak, competitive inhibitor with respect to GTP, IC50: 0.225 mM in the decarboxylation direction, modifications at N-1 and C-8 improve the inhibitory activity by more than 100fold
0.02
3-Mercaptopicolinic acid
Homo sapiens
-
reversible, non-competitive inhibitor, IC50: in the 0.02 mM range
0.3
KHCO3
Homo sapiens
-
-
9.3
L-alanine
Crassostrea gigas
Q0KHB7
in 100 mM imidazole-HCl buffer (pH 6.6), at 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.06
mutant enzyme D78A, phosphoenolpyruvate-forming activity in the presence of 0.2 mM Mn2+
0.13
mutant enzyme D78A, oxaloacetate-forming activity in the presence of 0.2 mM Mn2+
0.15
mutant enzyme D78A, oxaloacetate-forming activity in the presence of 2 mM Mn2+
0.36
mutant enzyme E83A, phosphoenolpyruvate-forming activity in the presence of 0.2 mM Mn2+
0.4
mutant enzyme E83A, oxaloacetate-forming activity in the presence of 0.2 mM Mn2+
1
mutant enzyme E83A, oxaloacetate-forming activity in the presence of 2 mM Mn2+
1.81
Modiolus demissus
-
-
2.93
-
-
4.14
-
in 50 mM Tris-HCl (pH 8.0) and 5 mM dithiothreitol, at 55C
4.75
-
-
10.6
mutant enzyme D75A, oxaloacetate-forming activity in the presence of 0.2 mM Mn2+
12.3
wild type enzyme, phosphoenolpyruvate-forming activity in the presence of 0.2 mM Mn2+
13.5
-
mitochondrial enzyme
14.5
-
cytosolic enzyme
15.5
mutant enzyme D75A, phosphoenolpyruvate-forming activity in the presence of 0.2 mM Mn2
16.7
-
carboxylation of phosphoenolpyruvate
19.9
-
decarboxylation of oxaloacetate
29.4
wild type enzyme, oxaloacetate-forming activity in the presence of 2 mM Mn2+
43
wild type enzyme, oxaloacetate-forming activity in the presence of 0.2 mM Mn2+
46.5
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2
-
phosphoenolpyruvate carboxylation and exchange reaction in presence of Mn2+
7.6
-
carboxylation, enzyme from heterotrophically grown cells
8.5
-
decarboxylation
8.8
-
decarboxylation, enzyme from autotrophically and heterotrophically grown cells
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.25 - 9
-
enzyme activity in the pH range of 6.25-9
6.3 - 7.5
Modiolus demissus
-
about 50% of maximal activity at pH 6.3 and pH 7.5
6.6 - 8
-
there is a steep rise in the enzyme activity (about 2fold) between the pH 6.6 and pH 7.4, although a sharp decline is observed within a narrow range (pH 7.48.0)
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35 - 40
-
-
37
-
assay at
additional information
-
assay at room temperature
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50 - 90
50C: about 45% of maximal activity, 90C: about 80% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5
-
isoelectric focusing
4.61
-
theoretically calculated pI
5.5
-
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
; PEPCK mRNAs and enzyme activities are measured in muscle during prolonged hypoxia for 20 days. The PEPCK mRNA ratio in hypoxic muscle significantly increases at day 10 simultaneously to the PEPCK enzyme activity
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25177 / H37Ra)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6925
deduced from cDNA
24000
-
all four enzyme forms, gel filtration
42000
-
mitochondrial enzyme, gel filtration
50000
-
gel filtration
56000
-
mitochondrial enzyme, gel filtration
60000
-
cytosolic enzyme, gel filtration
67000
-
-
67310
mature protein, calculated from amino acid sequence
67540
-
recombinant enzyme, mass spectrometry
69250
calculated from amino acid sequence
70000
Modiolus demissus
-
gel filtration
71200
calculated from the nucleic acid sequence-derived amino acid sequence
74000
-
gel filtration
75400
-
high speed equilibrium sedimentation
80000
-
non-denaturing PAGE
83000
-
gel filtration
83200
gel filtration
85000
-
gel filtration
284000
gel filtration
550000
-
enzyme from heterotrophically grown cells, gel filtration
761000
-
enzyme from autotrophically grown cells, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homotetramer
4 * 70000, SDS-PAGE; 4 * 72039, calculated from sequence
monomer
tetramer
trimer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method. Crystallizes in space group P21 with four molecules per asymmetric unit. The 2.3 A resolution structure is solved by molecular replacement using the human cytosolic PCK
-
X-ray analysis, hanging-drop method
-
hanging drop vapour diffusion method using 0.1 M HEPES (pH 7.4) and 19% PEG 6000
enzyme alone and in complexes with the non-hydrolyzable GTP analog beta,gamma-methylene GTP, and with phosphoenolpyruvate, hanging-drop vapor-diffusion method, X-ray analysis
-
hanging drop vapour diffusion method, at 25 ?C in 0.1 M HEPES (pH 7.4) and 12-30% PEG 3350, creating PEPCK-Mn2+, PEPCK-Mn2+-oxaloacetate, PEPCK-Mn2+-oxaloacetate-Mn2+GDP, and PEPCKMn2+-Mn2+GTP crystals
-
mutant enzyme A467G in complex with Mn2+, substrates and inhibitors
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
protein concentration 0.057 mg/ml, glutathione concentration 0.2 mM, stable for at least 3 h
80
half life: 53 min; half-life: 53 min
additional information
-
cytosolic enzyme is less stable against heat than mitochondrial enzyme
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol facilitates loss of activity
-
dithiothreitol stabilizes
-
photoinactivation by 8-azidoguanosine 5'-triphosphate
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
PEPCK is very stable toward oxidative treatment
-
649936
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, mitochondrial enzyme, stable for several months
-
-20C, 0.2 M phosphate, DTT and glycerol, stable for 1 month
-
-70C, 0.05 M potassium phosphate, pH 6.8, 0.2 M sucrose, stable for at least 21 months
-
-80C, more than 1 year, the enzyme remains stable
-
4C, 3 weeks, stable
-
4C, Co3+-PEPCK with Co3+ bound to enzyme at site n1, 1 week, stable
-
4C, Cr3+-PEPCK with Cr3+ bound to enzyme at site n1, at least 3 days, stable
-
4C, in absence of metal ions, under N2, protein concentration 0.5 mg/ml or higher, stable for at least 14 days
-
4C, purified recombinant enzyme, 100 mM sodium phosphate buffer, pH 7, 100 mM NaCl, 1 month, 24% loss of activity
4C, purified recombinant His-tagged enzyme, 100 mM sodium phosphate buffer, pH 7, 100 mM NaCl, 2 months, 25% loss of activity
room temperature, at least 6 h, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
7.5fold, recombinant enzyme
-
; recombinant protein
ammonium sulfate precipitation, DEAE-Sephacel column chromatography, and Blue Sepharose CL 6B affinity chromatography
-
cytoslic and mitochondrial enzyme
-
cytosolic enzyme
-
from autotrophically grown cells and heterotrophically grown cells
-
glutathione-Uniflow resin column chromatography and P6DG column chromatography
-
mutant PEP carboxykinase
Ni-NTA Superflow agarose column chromatography and Ekapture-agarose column chromatography
-
recombinant and mutant enzyme
-
recombinant enzyme
recombinant wild-type and mutant enzyme
-
Superflow Ni2+-nitrilotriacetic acid-agarose chromatography
-