Information on EC 2.7.9.1 - pyruvate, phosphate dikinase

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

EC NUMBER
COMMENTARY hide
2.7.9.1
-
RECOMMENDED NAME
GeneOntology No.
pyruvate, phosphate dikinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + pyruvate + phosphate = AMP + phosphoenolpyruvate + diphosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
C4 and CAM-carbon fixation
-
-
C4 photosynthetic carbon assimilation cycle, NAD-ME type
-
-
C4 photosynthetic carbon assimilation cycle, NADP-ME type
-
-
C4 photosynthetic carbon assimilation cycle, PEPCK type
-
-
Carbon fixation in photosynthetic organisms
-
-
Carbon fixation pathways in prokaryotes
-
-
L-glutamine biosynthesis III
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
photosynthesis
-
-
Pyruvate metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:pyruvate, phosphate phosphotransferase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9027-40-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Arundinaria sp.
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
anendosymbiont, Wolbachia sp. subsp. Brugia malayi
-
-
Manually annotated by BRENDA team
Crithidia sp.
-
-
-
Manually annotated by BRENDA team
barnyard grass
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Kalanchoe hildebrandtii
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Euryarchaea, Archaeabacteria
SwissProt
Manually annotated by BRENDA team
a bioenergy feedstock grass
-
-
Manually annotated by BRENDA team
Nopalxochia ackermannii
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Crenarchaea, Archaeabacteria
SwissProt
Manually annotated by BRENDA team
strain S-1
-
-
Manually annotated by BRENDA team
strain S-1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Sedum prealtum
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Rhizobium
-
-
Manually annotated by BRENDA team
spinach
-
-
Manually annotated by BRENDA team
the entire gene consists of 2748 bp, GC content of 52.6%, protein of 915 amino acids corresponding to a theoretical molecular mass of 102.3 kDa, sequence signatures compared to other PPDK genes; hyperthermophilic crenarchaeote, Archaeabacteria, mass culture of Thermoproteus strain DSM 2078
SwissProt
Manually annotated by BRENDA team
fragment; parabasalia, isolated from hindgut of damp-wood termites
SwissProt
Manually annotated by BRENDA team
cv. Lerma Rojo
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
-
the enzyme plays a controlling role in the phosphoenolpyruvate-regeneration phase of the C4 photosynthetic pathway
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',3'-dideoxyadenosine 5'-triphosphate + pyruvate + phosphate
?
show the reaction diagram
-
-
-
-
r
2'-dATP + pyruvate + phosphate
?
show the reaction diagram
-
-
-
-
r
3'-dATP + pyruvate + phosphate
?
show the reaction diagram
-
-
-
-
r
AMP + phosphoenolpyruvate + diphosphate
ATP + pyruvate + phosphate
show the reaction diagram
ATP + PPDK central domain construct of residues 381-512 (Cent-I) + phosphate
?
show the reaction diagram
-
-
-
?
ATP + PPDK N-terminal domain construct of residues 1-340 (Tem-340) + phosphate
?
show the reaction diagram
-
-
-
?
ATP + PPDK N-terminal domain construct of residues 1-553 (Tem340-Cent-I) + phosphate
?
show the reaction diagram
-
-
-
?
ATP + pyruvate + arsenate
?
show the reaction diagram
ATP + pyruvate + phosphate
AMP + phosphoenolpyruvate + 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
AMP + phosphoenolpyruvate + diphosphate
ATP + pyruvate + phosphate
show the reaction diagram
ATP + pyruvate + phosphate
AMP + phosphoenolpyruvate + diphosphate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Na+
-
slight activation of the phosphoenol pyruvate formation
Ni2+
-
1 mM of this ion promotes pyruvate formation
Rb+
-
25% of the activity relative to NH4+
Tl+
-
94% of the activity relative to NH4+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1-benzenesulfonyl-pyrrolidin-2-yl)-(3,5-dimethyl-4-p-tolylsulfanyl-pyrazol-1-yl)-methanone
-
(7-benzyl-3-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-ylsulfanyl)-acetic acid benzyl ester
-
2-(1-benzyl-1H-benzoimidazol-2-ylmethylsulfanyl)-3H-quinazolin-4-one
-
2-(1H-Benzoimidazol-2-ylsulfanyl)-N-(5-phenyl-[1,3,4]thiadiazol-2-yl)-acetamide
-
2-(1H-benzoimidazol-2-ylsulfanyl)-N-[4-(pyridin-2-ylsulfamoyl)-phenyl]-acetamide
-
2-(9-allyl-9H-1,3,4,9-tetraaza-fluoren-2-ylsulfanyl)-N-(5-methyl-isoxazol-3-yl)-butyramide
-
2-(9-benzyl-9H-1,3,4,9-tetraaza-fluoren-2-ylsulfanyl)-N-furan-2-ylmethyl-acetamide
-
2-benzylsulfanyl-N-[5-(3,4-dichloro-benzyl)-[1,3,4]thiadiazol-2-yl]-acetamide
-
2-Bromobutyrate
-
80% inhibition at 1 mM
2-Bromopropionate
-
74% inhibition at 1 mM
2-[2-(1-benzyl-1H-benzoimidazol-2-ylsulfanylmethyl)-benzoimidazol-1-yl]-acetamide
-
3-(2,6-dichloro-phenyl)-5-methyl-isoxazole-4-carboxylic acid 4-(6-amino-5-cyano-3-methyl-1,4-dihydro-pyrano[2,3-c]pyrazol-4-yl)-phenyl ester
-
3-Bromopropionate
-
66% inhibition at 1 mM
4-chloro-N-[2-(1-phenyl-1H-tetrazol-5-ylsulfanyl)-acenaphthen-1-yl]-benzenesulfonamide
-
4-[4-hydroxy-5-oxo-2-(3-phenoxy-phenyl)-1-pyridin-3-ylmethyl-2,5-dihydro-1H-pyrrole-3-carbonyl]-N,N-dimethyl-benzenesulfonamide
-
5'-adenylimidodiphosphate
-
-
5,5-dithiobis(2-nitrobenzoic acid)
5-benzyl-2-(4-chloro-phenyl)-3-(4-fluoro-phenyl)-tetrahydro-pyrrolo[3,4-d]isoxazole-4,6-dione
-
5-phenyl-2-[2-([1,2,4]triazolo[4,3-a]pyridin-3-ylsulfanyl)-acetylamino]-thiophene-3-carboxylic acid ethyl ester
-
6-(4-benzyl-5-phenyl-4H-[1,2,4]triazol-3-ylsulfanylmethyl)-N-phenyl-[1,3,5]triazine-2,4-diamine
-
6-[5-(2-chloro-phenyl)-4-phenyl-4H-[1,2,4]triazol-3-ylsulfanylmethyl]-N-phenyl-[1,3,5] triazine-2,4-diamine
-
adenosine 5'-mono-O-phosphorothiolate
-
-
alpha-beta-methylene ATP
beta-gamma-methylene ATP
biphosphonate
-
mixed inhibition mechanism with respect to diphosphate
Bromoacetate
-
83% inhibition at 1 mM
Bromopyruvate
Co2+
-
at high concentration inhibits the phosphoenolpyruvate, pyruvate exchange reaction
D-glucose 1-phosphate
72% activity sustained, effector concentration 5 mM, 55C
diethyldicarbonate
-
inactivates the enzyme by combination with histidyl residues, inhibition is reversed by hydroxylamine
diphosphate
gamma(p-Arsenophenyl)-n-butyrate
ilimaquinone
Imidodiphosphate
-
competitive to diphosphate
iodoacetate
Methylene diphosphonate
MgHPO4
-
competitive to HPO42-
Mn2+
-
at high concentration inhibits the phosphate, diphosphate exchange reaction
N-(3-cyano-4,5-diphenyl-furan-2-yl)-2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-acetamide
-
N-(3-cyano-4,5-diphenyl-furan-2-yl)-4-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-butyramide
-
N-ethylmaleimide
N-[5-(3-chloro-benzyl)-thiazol-2-yl]-3-[5-(2-methyl-cyclopropyl)-furan-2-yl]-propionamide
-
oxalate
p-chloromercuribenzoate
p-hydroxymercuribenzoate
phosphate
phosphoenolpyruvate
Phosphoglycolate
-
competitive to phosphoenolpyruvate
potassium fluoride
-
inhibits reaction in both directions at 50 mM
PPDK regulatory protein
-
RP catalyzes reversible phosphorylation on T456 by RP, inactive form phoshorylated at T456, ADP-dependent, a catalytic His-phophorylation precedes phosphorylation at T456, pyruvate (2 mM) can inhibit inactivation by RP
-
pyruvate
Sulfhydryl agents
tetrasodium 1-hydroxyethylidene biphosphonate
-
-
tetrasodium 1-hydroxymethylidene biphosphonate
-
-
tetrasodium 1-hydroxynonane biphosphonate
-
-
UDP
55% activity of PPDK, effector concentration 5 mM, 55C
unguinol
-
from fungal isolate F3000054, a mixed noncompetitive inhibitor of PPDK with respect to the substrates pyruvate and ATP and an uncompetitive inhibitor of PPDK with respect to phosphate. Unguinol has deleterious effects on a model C4 plant but no effect on a model C3 plant, effects in vivo, overview; mixed noncompetitive inhibition of PPDK with respect to the substrates pyruvate and ATP, uncompetitive with respect to phosphate, phytotoxic effects (bleaching) on C4 plants, no effect on a model C3 plant (Hordeum vulgare)
[3-methyl-7-(3-methyl-benzyl)-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-8-ylsulfanyl]-acetic acid benzyl ester
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
abscisic acid
-
induces production of the protein
mannitol
-
induces production of the protein
Polyethylene glycol
-
induces production of the protein
thiols
-
required for activity in solution
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.28
2',3'-dideoxyadenosine 5'-triphosphate
-
pH 7, 25C
0.1
2'-dAMP
-
pH 6.8, 25C
0.35
2'-dATP
-
pH 7, 25C
0.25
3'-dATP
-
pH 7, 25C
0.0013 - 0.02
AMP
0.004 - 8
ATP
0.029 - 3.5
diphosphate
0.0014 - 1.8
phosphate
0.02 - 0.5
phosphoenolpyruvate
0.025 - 305
pyruvate
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
53 - 87
phosphoenolpyruvate
10 - 15
pyruvate
additional information
additional information
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.07
5'-adenylimidodiphosphate
-
pH 6.8, 25C
0.32
adenosine
-
pH 6.8, 25C
0.31
adenosine 5'-mono-O-phosphorothiolate
-
pH 6.8, 25C
0.13 - 0.32
AMP
0.036 - 17
ATP
0.32 - 0.55
diphosphate
8.7
GMP
-
pH 6.8, 25C
0.001 - 0.025
oxalate
6 - 88
phosphate
0.15 - 2
phosphoenolpyruvate
0.4 - 22.6
pyruvate
1.2
tetrasodium 1-hydroxyethylidene biphosphonate
-
pH 6.3, 25C
11
tetrasodium 1-hydroxymethylidene biphosphonate
-
pH 6.3, 25C
4.3
tetrasodium 1-hydroxynonane biphosphonate
-
pH 6.3, 25C
additional information
additional information
-
inhibition kinetics for unguinol on PPDK
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.29 - 0.292
ilimaquinone
0.04
unguinol
Zea mays
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.043
-
T456F mutant
0.067
-
inactive enzyme obtained from dark-grown plants
0.24
-
T456Y mutant
1.2
-
measured at 30C
2.5
catalysis of both the pyruvate and the phosphoenolpyruvate formation
2.91
-
after activation by phosphate of an enzyme obtained from dark-grown plants
4.1
-
-
4.5
-
-
12
-
recombinant enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8 - 6.4
-
reaction with AMP, phosphoenolpyruvate and diphosphate
6
-
ATP formation
6.4 - 6.7
-
pyruvate formation
6.4
-
reverse reaction, pyruvate formation
6.5
-
pyruvate formation
6.5 - 7
-
both directions of reaction
6.5 - 7.5
-
phosphoenolpyruvate formation
6.8 - 7
-
assay at
6.9
-
pyruvate formation
7 - 7.8
-
phosphoenolpyruvate formation
7.1
-
pyruvate formation
7.2 - 7.8
-
forward reaction, phosphoenolpyruvate formation
7.2 - 8
-
forward reaction, phosphoenolpyruvate formation
7.75
-
pyrosequencing by utilizing pyruvate phosphate dikinase, expressed onto bacterial magnetic particles, PPDK-BacMPs
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 9.5
-
pyruvate phosphate dikinase, expressed onto bacterial magnetic particles, PPDK-BacMPs, shows stable enzymatic activity
7 - 8.3
-
the competency of the enzyme in catalyzing its phosphoenolpyruvate-forming reaction at pH 7.0 is dramatically reduced, having only 6% of the rate at pH 8.3
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at
37
-
activity assay; pyrosequencing by utilizing pyruvate phosphate dikinase, expressed onto bacterial magnetic particles, PPDK-BacMPs
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 45
-
pyruvate phosphate dikinase, expressed onto bacterial magnetic particles, PPDK-BacMPs, shows stable enzymatic activity
additional information
-
cold tolerance of recombinant maize plants estimated, crude leaf extracts placed on ice to measure residual PPDK activities, PPDK extracted from untransformed maize looses 90% of its activity after 20 min, a retained PPDK activity 70% of after 180 min in two transformants corresponds to a cold-tolerance shown for Flaveria brownii plants
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
isoelectric focusing (pH range from 4 to 7), cytosolic CyPPDK 2
5.3
isoelectric focusing (pH range from 4 to 7), cytosolic CyPPDK 2
5.4
isoelectric focusing (pH range from 4 to 7), cytosolic CyPPDK 2
8.25
calculated
8.9
-
calculated
additional information
annotation of different isoforms of one of the two cytosolic isoforms of PPDK of maize
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
cytosolic mRNA of OsPPDKB is induced in the reproductive organs after pollination, and greatly increases until about 10 days after fertilization. This mRNA is localized mainly in the endosperm, aleurone, and scutellum of the developing kernel
Manually annotated by BRENDA team
-
the cytosolic isoform accumulates preferentially in the veins
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
cytosolic mRNA of OsPPDKB is induced in the reproductive organs after pollination, and greatly increases until about 10 days after fertilization. This mRNA is localized mainly in the endosperm, aleurone, and scutellum of the developing kernel
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
96300
calculated
96600
-
calculated
99280
cytosolic CyPPDK 2
100000
102300
theoretical weight calculated from sequence
104900
cytosolic CyPPDK 2
105100
cytosolic CyPPDK 2
105700
cytosolic CyPPDK 2
150000 - 160000
-
equilibrium sedimentation and gel filtration
150000
169300
-
sucrose density gradient sedimentation
170000
-
gel filtration
195000
-
gel filtration
198400
-
gel electrophoresis
246800
-
HPLC gel filtration
250000
native protein, gelfiltration
320000
-
gel filtration on Sephadex G-200
330000
-
gel filtration on Sephrose 6B
370000
385000
-
gel filtration
387000
-
sedimentation analysis
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
-
6 * 58000, SDS-PAGE
homodimer
homotetramer
-
4 * 95000, the enzyme is maximally active as a homotetramer and is inactive in the dimeric and monomeric forms
monomer
-
1 * 13438, recombinant protein of the central domain of PPDK (Cent-I protein), mass spectrometry, chromatography
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
additional information
-
during seed development posttranslational down-regulation of PPDK in activity and amount by regulatory threonyl-phosphorylation PPDK regulatory protein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
method description
-
PPDK, mutant R219E/E271R/S262D, hanging drop vapour diffusion method at 30C, using a protein solution containing 28 mg/ml protein in 20 mM imidazole, pH 6.5, 0.1 mM EDTA, 100 mM KCl, and 1 mM DTT. The reservoir solution contains 50% saturated ammonium sulfate and 0.1 M Na-HEPES, pH 7.0, mixing of equal volumes of protein solution and reservoir solution, structure determination and analysis of two different enzyme conformations at 1.94 A resolution, modeling
-
X-ray crystal structure mentioned, the central domain faces the C-terminal domain and positions the phosphorylated His residue for phosphoryl transfer to pyruvate thereby forming phosphoenolpyruvate and regenerating the unphosphorylated H455, native structure determined by NMR
-
crystals sensitive to temperature
-
hanging-drop vapour diffusion method, crystal strcuture with and without phosphoenolpyruvate, determined at 2.3 A resolution
-
method description
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2
-
activity in vivo decreases more than 30% when exposed 90 min
12
-
loses activity below at about 12C by dissociation of the tetramer, considered as one possible cause of the reduction of the photosynthetic rate of maize at low temperatures
20 - 40
-
stable for at least 30 min
24
-
45 min, 0.6 mg protein/ml, 18% loss of activity. 2.4 mg protein/ml, no inhibition
50
-
irreversible denaturation
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
central domain protein Cent-I, 4.3 kcal/mol (delta G) for unfolding at 25C in buffer at pH 7.0
-
does not require thiol compounds to maintain stability during storage or assay
-
freezing and thawing inactivates
-
glycerol protects both the day-form and night-form in vitro
-
Mg2+ stabilizes the oligomeric structure of the enzyme
-
sensitive to dilution, particularly at concentrations below 0.3 mg/ml, stable to freezing and thawing
-
slowly inactivates when kept at 4C
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50% v/v glycerol, optimal storage condition, decay factor is 3 months
-
0C, as a precipitate in a 66% saturated solution of (NH4)2SO4
22C, Tris-HCl buffer, pH 7.9, 5 mM imidazole, 50 mM NaCl, 2 weeks
-
retains 85% of activity after two weeks at room temperature
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
isolation of an inactive enzyme form dark-grown leaves
-
maize PPDK of leaves expressed for inhibitor studies in Escherichia coli with a His-tag and purified on a Ni-affinity column
-
method that includes metal affinity chromatography
-
method that includes successive chromatography through DE-52, hydroxyapatite, Sephadex G-200 and Blue agarose
-
partial
partial by a method that includes Sephadex G-25 chromatography
-
partial, using ammonium sulfate precipitation and chromatography on DEAE-cellulose
-
partial, using ammonium sulfate precipitation and chromatography on DEAE-cellulose and hydroxyapatite
-
partial, using ammonium sulfate precipitation and chromatography on DEAE-cellulose and Sephadex G-200
-
partial, using ammonium sulfate precipitation, and chromatography on DEAE-cellulose and Bio-Gel
-
partial, using Sephadex G-25 and DEAE column chromatography
partial, using Sephadex G-25 column chromatography
partial, using Sephadex-G200 and Hypatite C chromatography
-
recombinant maize PPDK used for inactivation/activation assay by the PPDK regulatory protein RP
-
recombinant protein, heat precipitation at 90C for 20 min, ion exchange chromatography
recombinant proteins of the central domain (Cent-I) and of the N-terminus (Tem-340, residues 1-340 of the native PPDK)
-
soluble recombinant His-tagged PPDK from Escherichia coli by nickel affinity chromatography
-
two deletion mutants purified by a method that includes DEAE-cellulose and Sephadex G-200 chromatography
-
using hydroxyapatite, Sephadex G-200 and DEAE column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
25 kDa C-terminal and 35 kDa N-terminal deletion mutants expressed in Escherichia coli
-
78% of homology with maize enzyme
-
; expressed in Escherichia coli
-
Escherichia coli, strains DH5alpha and BL21-CodonPLUS(DE3)-RIL
Escherichia coli, three constructs, one construct consisting of the 3'-part of Flaveria brownii (Asteraceae) of cold tolerant PPDK fused to maize PPDK (15th exon), another construct includes a set of point mutations to substitute all of the 17 residues that differ between the 3'-parts of maize and Flaveria brownii PPDK, respectively, the whole genomic sequence of the maize PPDK gene is included as a control, transformation of constructs into maize inbred line A188 by Agrobacterium tumefaciens (strain LBA4404), individual range of variation in the amount of PPDK among regenerated plants, crude leaf extracts of some transformed plants produce a large amount of cold tolerant recombinant enzyme and reveal a greatly improved cold tolerance especially by using the construct altered at 17 amino acid positions
-
Escherichia coli; Escherichia coli
expressed in a baculovirus system
-
expressed in Arabidopsis thaliana, found exclusively in chloroplasts of transgenic Arabidopsis plants
-
expressed in Escherichia coli
expressed in Nicotiana tabacum
-
expressed in Oryza sativa subsp. indica cultivar IR64
-
expression of mutant enzyme R219E/E271R/S262D in Escherichia coli strain JM101
-
genomic analysis of the glycolytic/gluconeogenic pathway, optimization of C-terminally His-tagged PPDK expression in Escherichia coli as soluble protein, overview
-
high levels of expression in Zea mays by using a double intron cassette and a chimeric cDNA made from Flaveria bidentis and Flaveria brownii with a maximum content of 1 mg/g fresh weight. In leaves of transgenic maize, PPDK molecules produced from the transgene are detected in cold-tolerant homotetramers or in heterotetramers of intermediate cold susceptibility formed with the internal PPDK
-
into the vector pUMP16M13
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introduction of a cold-tolerant PPDK cDNA isolated from Flaveria brownii into maize by Agrobacterium-mediated transformation. Higher levels of expression by using a double intron cassette and a chimeric cDNA made from Flaveria bidentis and Flaveria brownii with a maximum content of 1 mg/g fresh weight. In leaves of transgenic maize, PPDK molecules produced from the transgene are detected in cold-tolerant homotetramers or in heterotetramers of intermediate cold susceptibility formed with the internal PPDK
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phylogenetic analysis
quantitative expression analysis of the enzyme during endosperm development, expression profiles, overview
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quantitative real-time reverse transcription (RT)-PCR expression analysis during transfer of plants from 25C to 14C growth temperature
the cloned sequence represents about 20% of the complete gene, and shows about 56% homology with enzymes from maize and Bacteroides symbiosus
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to improve the cold stability of the enzyme, a cold-tolerant PPDK cDNA isolated from Flaveria brownii is introduced into maize by Agrobacterium-mediated transformation. Higher levels of expression are ontained by using a double intron cassette and a chimeric cDNA made from Flaveria bidentis and Flaveria brownii with a maximum content of 1 mg/g fresh weight. In leaves of transgenic maize, PPDK molecules produced from the transgene are detected in cold-tolerant homotetramers or in heterotetramers of intermediate cold susceptibility formed with the internal PPDK. A significant improvement in the cold stability of PPDK can be achieved when a suffcient quantity of cold-tolerant subunits is expressed in transgenic maize leaves
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
both the cytosolic and chloroplastic isoforms of pyruvate, orthophosphate dikinase are up-regulated in naturally senescing leaves
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in the C4 pathway, enzyme activity is strictly regulated in an up/down manner by the level of incident light
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D280A
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partial activity
E279A
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partial activity
R135A
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partial activity, 15 mutant enzymes studied
R219E/E271R/S262D
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site-directed mutagensis, comparison of mutant to wild-type enzyme structure
H458N
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no activity
T456D
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no activity
T456E
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no activity
T456S
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111% activity with respect to wild type
T456V
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98% activity with respect to wild type
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
percent decrease of fluorescence intensity (304 nm) for conversion of native protein (0 M urea) to fully denaturation (6 M urea) calculated (23.5%), DELTA G for unfolding of Cent-I (PPDK central domain construct of residues 381-512) in solvent determined (4.3 kcal/mol)
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
analysis
drug development
pharmacology
drug design, in silico studies on stereo chemical quality of PPDK protein structure, interaction studies to identify promising ligands to inhibit the function of PPDK, possibility of using proposed ligands as inhibitors for intestinal infections caused by Entamoeba histolytica in humans and for related pathogens, virtual screening of ligands to inhibit PPDK by docking studies using compound input libraries, phylogenetic trees of pathogens as further targets for in silico drug design to inhibit PPDK
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