Information on EC 1.1.1.95 - phosphoglycerate dehydrogenase

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

EC NUMBER
COMMENTARY hide
1.1.1.95
-
RECOMMENDED NAME
GeneOntology No.
phosphoglycerate dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3-phospho-D-glycerate + NAD+ = 3-phosphonooxypyruvate + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Glycine, serine and threonine metabolism
-
-
L-serine biosynthesis
-
-
Metabolic pathways
-
-
Methane metabolism
-
-
Microbial metabolism in diverse environments
-
-
serine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
3-phospho-D-glycerate:NAD+ 2-oxidoreductase
This enzyme catalyses the first committed step in the phosphoserine pathway of serine biosynthesis in Escherichia coli [2,3]. Reaction (1) occurs predominantly in the reverse direction and is inhibited by serine and glycine. The enzyme is unusual in that it also acts as a D- and L-2-hydroxyglutarate dehydrogenase (with the D-form being the better substrate) and as a 2-oxoglutarate reductase [3]. It has been postulated [3] that the cellular 2-oxoglutarate concentration may regulate serine biosynthesis and one-carbon metabolism directly by modulating the activity of this enzyme.
CAS REGISTRY NUMBER
COMMENTARY hide
9075-29-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene pgk
-
-
Manually annotated by BRENDA team
Frog
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Nitrosolobus sp.
gene pgk, several strains, overview
-
-
Manually annotated by BRENDA team
gene pgk, several strains, overview
-
-
Manually annotated by BRENDA team
gene pgk or cbbK, strain ATCC 25196
SwissProt
Manually annotated by BRENDA team
gene pgk, several strains, overview
-
-
Manually annotated by BRENDA team
Nitrosovibrio sp.
gene pgk, several strains, overview
-
-
Manually annotated by BRENDA team
Pigeon
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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
2-oxoglutarate + NADH
2-hydroxyglutarate + NAD+
show the reaction diagram
-
-
-
-
?
2-oxoglutarate + NADH + H+
2-hydroxyglutarate + NAD+
show the reaction diagram
-
-
-
-
?
2-phospho-D-glycerate + NAD+
2-phosphohydroxypyruvate + NADH + H+
show the reaction diagram
-
activity relative to 3-phospho-D-glycerate: 47%
-
-
?
3-phospho-D-glycerate + NAD+
3-phosphohydroxypyruvate + NADH
show the reaction diagram
3-phospho-D-glycerate + NAD+
3-phosphohydroxypyruvate + NADH + H+
show the reaction diagram
3-phospho-D-glycerate + NADP+
3-phosphohydroxypyruvate + NADPH + H+
show the reaction diagram
3-phosphoglycerate + NAD+
3-phosphohydroxypyruvate + NADH
show the reaction diagram
3-phosphohydroxypyruvate + NAD+
?
show the reaction diagram
-
-
-
r
3-phosphohydroxypyruvate + NADH
3-phosphoglycerate + NAD+
show the reaction diagram
-
specific for
-
-
?
alpha-ketoglutarate + NADH
2-hydroxyglutaric acid + NAD+
show the reaction diagram
DL-glyceraldehyde 3-phosphate + NAD+
?
show the reaction diagram
-
activity relative to 3-phospho-D-glycerate: 9%
-
-
?
phosphonooxypyruvate + NADH + H+
?
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
3-phospho-D-glycerate + NAD+
3-phosphohydroxypyruvate + NADH
show the reaction diagram
3-phosphoglycerate + NAD+
3-phosphohydroxypyruvate + NADH
show the reaction diagram
phosphonooxypyruvate + NADH + H+
?
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-acetylpyridine-NAD+
3-acetylpyridine-NADH
deamino-NAD+
deamino-NADH
-
as effective as NADH
thio-NAD+
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
optimal salt concentration is 350-400 mM
NaCl
optimal salt concentration is 350-400 mM
potassium phosphate
-
optimal activity at 75-100 mM, only about 30% of the optimal activity in 5 mM potassium phosphate
SO42-
-
stimulates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2SO4
-
inhibitory effect on NADPH-dependent 3-phosphohydroxypyruvate reduction
(R)-2-amino-1-propanol
-
slightly
3-phosphoglycerate
3-phosphohydroxypyruvate
Ag+
-
inhibition can be overcome by addition of dithiothreitol
AMP
-
weak inhibitory effect on 3-phospho-D-glycerate oxidation, remaining activity: 75%
beta-Alanine
-
slightly
glycine
hydroxyglutarate
-
product inhibition of the alpha-ketoglutarate reduction
iodoacetate
-
-
K2HPO4
-
inhibitory effect on NADPH-dependent 3-phosphohydroxypyruvate reduction. It appears that the phosphate ion PO43- exerts its inhibitory effect by binding to the free enzyme and NADPH-enzyme complex
L-alanine
L-allothreonine
-
-
L-homoserine
-
-
L-serine
L-threonine
-
-
Mercurials
-
inhibition can be overcome by addition of dithiothreitol
-
N-Butylmaleimide
-
-
N-ethylmaleimide
N-Heptylmaleimide
-
-
N-Pentylmaleimide
-
-
N-Phenylmaleimide
-
-
NAD+
-
product inhibition, competitive to NADH
p-chloromercuribenzoate
-
-
p-hydroxymercuribenzoate
-
-
pyridoxal 5'-phosphate
-
-
pyridoxamine 5'-phosphate
-
-
sulfhydryl reagents
-
-
ZnCl2
-
inhibitory effect on 3-phosphohydroxypyruvate reduction, remaining activity: 29%
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(NH4)2SO4
-
enhancing effect on NAD-dependent 3-phospho-D-glycerate oxidation and 3-phosphohydroxypyruvate reduction but inhibitory effect on NADPH-dependent 3-phosphohydroxypyruvate reduction
HPO42-
-
-
K2HPO4
-
enhancing effect on NAD-dependent 3-phospho-D-glycerate oxidation and 3-phosphohydroxypyruvate reduction but inhibitory effect on NADPH-dependent 3-phosphohydroxypyruvate reduction
L-methionine
-
maximum activation at 10 mM
NaCl
-
PGDH activity increase about 1.5-2.3fold upon the increase of salinity from 0.5 to 2.5 M NaCl
SO42-
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00025 - 0.0054
3-acetylpyridine-NAD+
0.00016
3-acetylpyridine-NADH
-
25C
0.033 - 320
3-phospho-D-glycerate
0.212
3-phosphoglycerate
pH 9.0
0.00013 - 40.2
3-phosphohydroxypyruvate
0.038 - 0.088
alpha-ketoglutarate
0.05 - 1.35
D-3-phosphoglycerate
0.0078 - 0.5
NAD+
0.0005 - 12
NADH
0.02 - 0.15
NADPH
0.015
phosphohydroxypyruvate
pH 6.5
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.2 - 18.7
3-phospho-D-glycerate
0.463 - 2461
3-phosphohydroxypyruvate
0.128 - 0.555
alpha-ketoglutarate
603 - 2827
NADH
additional information
additional information
Escherichia coli
-
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.054 - 7.218
3-phosphohydroxypyruvate
2 - 10
K2HPO4
0.22 - 2.32
L-serine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.14
-
3-phosphoglycerate oxidation, recombinant protein
8.7
-
3-phosphohydroxypyruvate reduction
9.7
-
alpha-ketoglutarate reduction
10.95
-
3-phosphohydroxypyruvate reduction, recombinant protein
21.3
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
enzyme possess a dual pH optimum. A significant decrease in the Ki for substrate inhibition at pH values corresponding to the valley between these optima is responsible for this phenomenon
5.5
-
NADH oxidation
6 - 6.5
reaction with 3-phosphohydroxypyruvate + NADH
6.1
-
NADH oxidation
7.1
assay at
8
-
enzyme possess a dual pH optimum. A significant decrease in the Ki for substrate inhibition at pH values corresponding to the valley between these optima is responsible for this phenomenon
9
reaction with 3-phosphoglycerate and NAD+
9.4
-
NAD+ reduction
10
-
3-phospho-D-glycerate oxidation
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.8
-
about 50% of activity maximum at pH 5 and 7.8, NADH oxidation
5.2 - 5.7
-
at approximately pH 5.7 the activity starts increasing again and reaches a new optimum at approximately pH 5.2 before decreasing once again
5.5 - 8.5
-
Tris buffer and phosphate buffer
6 - 8.5
-
activity drops sharply above pH 8.5
6.5 - 8
-
enzymatic activity decreases below pH 6.5
7.5
-
3-phosphohydroxypyruvate reduction
7.5 - 9.5
-
activity is relatively constant between pH 7.5-9.5. Above pH 9.5 and below pH 7.5 activity falls off rapidly
8.5 - 10.3
-
about 50% of activity maximum at pH 8.5 and 10.3, NAD+ reduction
9
-
3-phospho-D-glycerate oxidation
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45 - 50
-
reduction, but enzyme is thermally unstable, therefore assay is performed at 37C
50
-
assay at
53
-
optimum temperature for 3-phospho-D-glycerate oxidation
80
-
optimum for 3-phosphohydroxypyruvate reduction
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.9
calculation from nucleotide sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
stellate cells
Manually annotated by BRENDA team
-
sustentacular cell
Manually annotated by BRENDA team
high expression levels in the proliferative phase
Manually annotated by BRENDA team
-
etiolated
Manually annotated by BRENDA team
skin-derived
Manually annotated by BRENDA team
-
satellite cell of the dorsal root ganglia and intestinal nerve plexuses
Manually annotated by BRENDA team
-
located in the rostral migratory stream, high enzyme expression level
Manually annotated by BRENDA team
-
light-grown more than dark-grown
Manually annotated by BRENDA team
-
epithelium
Manually annotated by BRENDA team
-
satellite cell of the dorsal root ganglia and intestinal nerve plexuses
Manually annotated by BRENDA team
-
primary culture
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
light-grown more than dark-grown
Manually annotated by BRENDA team
-
of the dorsal root ganglia and intestinal nerve plexuses
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Brucella abortus (strain 2308)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Ralstonia solanacearum (strain GMI1000)
Sulfolobus tokodaii (strain DSM 16993 / JCM 10545 / NBRC 100140 / 7)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
56410
-
deduced from cDNA
57000
-
SDS-PAGE
70000 - 74000
gel filtration
70000
-
homodimer, gel filtration, pH 7
163000
-
sedimentation equilibrium analysis
165000
166000
-
gel filtration
247000
-
HPLC gel filtration
250000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
homotetramer
4 * 56800
monomer
-
1 * 35000 at pH 5.0
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant selenomethionine-labeled enzyme, in presence of 2 mM NAD+ and 5 mM 2-ketoglutarate, euqilibration against precipitation solution containing 1.18-1.25 M ammonium sulfate and 100 mM potassium phosphate, pH 6.4, 18C, 1-2 weeks, X-ray diffraction structure determination and analysis at 2.24 A resolution
-
D-3-phosphoglycerate dehydrogenase with bound effector L-serine or with bound substrate hydroxypyruvic acid phosphate, PGDH at 10 mg/ml is mixed with 5 mM hydroxypyruvic acid phosphate and 5 mM NAD+ analogue 3-acetyl pyridine adenine dinucleotide, or with 5 mM NADH and 5 mM L-serine, from 1 M Na K tartrate, 0.1 M MES, pH 6.5, cryoprotection in 25% propylene glycol, X-ray diffraction structure determination and analysis at resolutions of 2.7 A and 2.4 A, respectively
-
vapor diffusion in hanging drops, structure refined to 2.3 A resolution using SeMet multiwavelength anamolous dispersion
-
crystal structures is determined using X-ray diffraction to resolution of 1.95 A, crystals are grown at room temperature by sitting-drop method
crystal structure is determined using X-ray diffraction to resolution of 1.77 A, crystals are grown at room temperature by sitting-drop method
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 11.5
-
enzyme is stable over a wide range of pHs. No decrease in activity after 20 min incubation at 50C
684698
5.5 - 7.5
-
stable
287525
8.5 - 9.5
-
unstable
287525
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
3 min, 40% loss of reductase activity
50 - 80
-
enzyme is highly thermostable, retaining more than 90% of its activity after incubation for 1 h at 80C
additional information
-
activity remains stable after incubation at 40C for 7 min, but decreases to 50% when the enzyme is incubated at 50C for 7 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
at low ionic strength the enzyme irreversibly loses activity with time. In 20 mM phosphate buffer, pH 7.5, most of the activity is lost within 24 h. The activity loss is prevented if the ionic strength is kept above approximately 100 mM salt
-
dilution inactivates
dithiothreitol increases stability
enzyme is highly thermostable
-
inorganic ions, i.e. Cl-, SO42-, HPO42-, essential for stability
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
purified protein is extremly susceptible to oxidation at alkaline pH
-
287544
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, stable for several weeks
-
-75C, 15% glycerol, purified stable for months
-
-80C, 20 mM sodium phosphate buffer, pH 7.2
-
-80C, 50% glycerol, purified recombinant enzyme remains active after 1 month of storage
2C, 40% loss of activity, 7 days
-
4C, purified stable for weeks
-
5C, 60% loss of activity, 24 h
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
diverse mutants expressed in Escherichia coli
-
partial
-
PGDH is expressed in Escherichia coli and purified using 5'-AMP-Sepharose affinity chromatography
-
recombinant selenomethionine-labeled enzyme from strain B834
-
recombinant tagged wild-type and mutant enzymes
-
using a butyl Sepharose 4 Fast Flow column and a Superdex 200 gel filtration column. During purification an unexpected decline in enzyme activity is observed if the enzyme is stored in plastic tubes, but not in glass ones
-
using ammonium sulphate fractionation and ion exchange chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
diverse mutants expressed in Escherichia coli
-
DNA and amino sequence analysis, phylogenetic analysis, sequence comparisons
-
DNA and amino sequence analysis, phylogenetic analysis, sequence comparisons, expression in Escherichia coli strain JM109
expressed in Escherichia coli
expressed in Escherichia coli and Arabidopsis thaliana
-
expression as selenomethionine-labeled enzyme in strain B834
-
expression in Escherichia coli
expression in Escherichia coli BL21
-
expression in Escheriochia coli BL21
expression of tagged wild-type and mutant enzymes
-
gene PHGDH consists of 12 exons and maps to chromosome 1p12, enzyme expression analysis in fibroblasts, transient overexpression of FLAG-tagged wild-type and mutant enzymes in HEK-293 cells
gene PHGDH, quantitative expression analysis in wild-type and mutant mice, genotyping, overview
-
gene serA, overexpression in Escherichia coli strains DH5alphaMCR and GM2929, co-overexpression of serA with other enzymes of the L-serine biosynthesis pathway leads to increased L-serine production in Corynebacterium glutamicum mutant serADELAT197
-
overexpression in Escherichia coli strain AD494 (DE3), cloning of GFP-mutant with transit peptide, expression in plant
-
overexpression of serA gene, that codes for the enzyme, in Escherichia coli strain JM105
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
HOXA10 is required for enzyme regulation in the endometrium, HOAX antisense expression, siRNA, causes a 4.4fold upregulation of 3-PGDH in endometrial cells
HOXA10 is required for enzyme regulation in the endometrium, HOAX sense expression causes a 2.0fold downregulation of 3-PGDH in endometrial cells
Phgdh knockout mouse embryos demonstrate that free serine and glycine concentrations are decreased markedly in head samples
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E108A
-
in contrast to wild-type mutant existed as monomer even at pH 7
A143A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
A144V
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
A374V
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
D317A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
D386A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
E299A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
E302A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
E307A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
E345A
-
site-directed mutagenesis, mutation of a residue in the serine binding site, the mutant shows only slightly altered kinetics and activity compared to the wild-type enzyme
E360A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
E387A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
G145V
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
G336V/G337V
-
changing glycine residues 336 and 337 to valine affect the sensitivity of the enzyme to inhibition by L-serine but not the extent of inhibition. The decrease in sensitivity is caused primarily by a decrease in the affinity of the enzyme for L-serine. The mutations also affect the domain rotation of the subunits in response to L-serine binding
G337V
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
G349V
-
site-directed mutagenesis, mutation of a residue in the serine binding site, the mutant shows only slightly altered kinetics and activity compared to the wild-type enzyme
G362V
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
H335A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
H344A/N364A
-
no inhibition by L-serine
K141A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
K311A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
N190A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, no protein expression
N303A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
N303A/K311A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
N346A/H344A
-
no inhibition by L-serine
N346A/N364A
-
no inhibition by L-serine
P348A
-
site-directed mutagenesis, mutation of a residue in the serine binding site, the mutant shows only slightly altered kinetics and activity compared to the wild-type enzyme
Q298A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
Q301A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
Q361A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
Q375A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
R338A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
R347A
-
site-directed mutagenesis, mutation of a residue in the serine binding site, the mutant shows only slightly altered kinetics and activity compared to the wild-type enzyme
S107A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S107A/S111A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S111A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S111A/K311A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S296A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S316A
-
site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S323A
-
site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
S373A
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site-directed mutagenesis, mutation of a residue in the Trp-139-loop and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
T297A
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site-directed mutagenesis, mutation of a residue in the polypeptide connecting the substrate binding domain and the regulatory domain, the mutant shows altered kinetics and reduced activity compared to the wild-type enzyme
W139F/E360W
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exhibits catalytic activity comparable to that of the native enzyme and is effectively inhibited by L-serine. The only fluorescence signal of the mutant is due to the single tryptophan at position 360. Pre-steady state analysis of binding of inhibitor serine shows that each serine binding interface produces an integrated fluorescent signal
W139F/E360W/G294V
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placement of a tryptophanyl residue near the serine binding site (W139F/E360W) allows serine binding to be monitored by fluorescence quenching analysis. Pre-steady state analysis demonstrate that serine binds to two forms of the free enzyme, E and E*. Conversion of Gly-336 to valine has its main effect on the Kd of serine binding to one form of the free enzyme (E) while maintaining the cooperativity of binding observed in the native enzyme
W139F/E360W/G336V
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placement of a tryptophanyl residue near the serine binding site (W139F/E360W) allows serine binding to be monitored by fluorescence quenching analysis. Pre-steady state analysis demonstrate that serine binds to two forms of the free enzyme, E and E*. Conversion of Gly-294 to valine eliminates a rate limiting conformational change that follows serine binding to E. The conformational change between the two forms of free enzyme is maintained, but the Hill coefficient for cooperativity is significantly lowered
A373T
naturally occuring mutation located in the nucleotide binding and regulatory domains of 3-PGDH, the mutation does not affect steady-state expression, protein stability, and protein degradation rates, the mutant is almost catalytically inactive
G377S
naturally occuring mutation located in the nucleotide binding and regulatory domains of 3-PGDH, the mutation does not affect steady-state expression, protein stability, and protein degradation rates, the mutant is almost catalytically inactive
R135W
naturally occuring mutation located in the nucleotide binding and regulatory domains of 3-PGDH, the mutation does not affect steady-state expression, protein stability, and protein degradation rates, the mutant is almost catalytically inactive
V261M
naturally occuring mutation located in the nucleotide binding and regulatory domains of 3-PGDH, the mutation does not affect steady-state expression, protein stability, and protein degradation rates, the mutant is almost catalytically inactive
V425M
naturally occuring mutation in the carboxy-terminal regulatory domain, leads to 3-PGDH deficiency, the mutant is almost catalytically inactive
V490M
naturally occuring mutation in the carboxy-terminal regulatory domain, leads to 3-PGDH deficiency, the mutant is almost catalytically inactive
up
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Phgdh is transiently upregulated in the brain by kainic acid-treatment, causing selective lesions in the hippocampal region, kainic acid is injected into the bregma, overview
D463A
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site-directed mutagenesis, a very large reduction in the sensitivity of the mutant enzyme to L-serine
G316V
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site-directed mutagenesis, the mutant shows slightly reduced activity and decreased sensitivity to L-serine compared to the wild-type
G316V/G317V
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site-directed mutagenesis, the mutant shows reduced activity and decreased sensitivity to L-serine compared to the wild-type
G316V/G317V/G318V
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site-directed mutagenesis, the mutant is not producable
G316V/G318V
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site-directed mutagenesis, the mutant shows reduced activity and decreased sensitivity to L-serine compared to the wild-type
G317V
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site-directed mutagenesis, the mutant shows slightly reduced activity and decreased sensitivity to L-serine compared to the wild-type
G317V/G318V
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site-directed mutagenesis, the mutant shows reduced activity and decreased sensitivity to L-serine compared to the wild-type
G318V
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site-directed mutagenesis, the mutant shows slightly reduced activity and decreased sensitivity to L-serine compared to the wild-type
K439A/R451A/R501A
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site-directed mutagenesis, the mutation eliminates substrate inhibition and pH-dependent depression in activity
N481A
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site-directed mutagenesis, a very large reduction in the sensitivity of the mutant enzyme to L-serine. Mutant N481A co-elutes with native PGDH in gel filtration, it shows loss of cooperativity, which cannot be explained by a change in the quaternary structure of the enzyme from tetramer to dimer or monomer
R451A/R501A/K439A
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site-directed mutagenesis, the mutation eliminates substrate inhibition and pH-dependent depression in activity
R501A/R451A/K439A
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anion binding site mutant: Km (mM) (3-phosphohydroxypyruvate): 0.243, kcat: 1558, Ki (mM) (3-phosphohydroxypyruvate): 7.218, mutant displays only little uncompetitive substrate inhibition, no dual pH optima compared to wild-type
W130F
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site-directed mutagenesis, catalytically inactive mutant
W29F
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
W327F
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
Y461A
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site-directed mutagenesis, a very large reduction in the sensitivity of the mutant enzyme to L-serine
H447A
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site-directed mutagenesis, the mutant shows altered L-serine binding, kinetics for NADH, and activity compared to the wild-type enzyme
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K439A
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site-directed mutagenesis, the mutant shows altered L-serine binding, kinetics for NADH, and activity compared to the wild-type enzyme
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R446A
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site-directed mutagenesis, the mutant shows altered L-serine binding, kinetics for NADH, and activity compared to the wild-type enzyme
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R451A
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site-directed mutagenesis, the mutant shows altered L-serine binding, kinetics for NADH, and activity compared to the wild-type enzyme
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R501A
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site-directed mutagenesis, the mutant shows altered L-serine binding, kinetics for NADH, and activity compared to the wild-type enzyme
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additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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metabolic engineering of Corynebacterium glutamicum for L-serine production by enzyme overexpression
molecular biology
synthesis
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metabolic engineering of Corynebacterium glutamicum for L-serine production by enzyme overexpression
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