Information on EC 1.1.1.8 - glycerol-3-phosphate dehydrogenase (NAD+)

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

EC NUMBER
COMMENTARY hide
1.1.1.8
-
RECOMMENDED NAME
GeneOntology No.
glycerol-3-phosphate dehydrogenase (NAD+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
sn-glycerol 3-phosphate + acceptor = glycerone phosphate + reduced acceptor
show the reaction diagram
sn-glycerol 3-phosphate + NAD+ = glycerone phosphate + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
1,3-propanediol biosynthesis (engineered)
-
-
Biosynthesis of secondary metabolites
-
-
glycerol-3-phosphate shuttle
-
-
Glycerophospholipid metabolism
-
-
superpathway of phosphatidate biosynthesis (yeast)
-
-
SYSTEMATIC NAME
IUBMB Comments
sn-glycerol-3-phosphate:NAD+ 2-oxidoreductase
Also acts on propane-1,2-diol phosphate and glycerone sulfate (but with a much lower affinity).
CAS REGISTRY NUMBER
COMMENTARY hide
9075-65-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
honey bee
-
-
Manually annotated by BRENDA team
bumblebee species
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain Wiedemann, mediterranean fruit fly
-
-
Manually annotated by BRENDA team
strain Wiedemann, mediterranean fruit fly
-
-
Manually annotated by BRENDA team
strain 11/32-90, green alga
-
-
Manually annotated by BRENDA team
three GPDH genes, CrGPDH1, CrGPDH2, and CrGPDH3
-
-
Manually annotated by BRENDA team
japanese quail
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
from Southeastern Wyoming
-
-
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
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
; jerboa
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain Lou/C
-
-
Manually annotated by BRENDA team
Rattus norvegicus Sprague-Dawley
Sprague-Dawley
-
-
Manually annotated by BRENDA team
strain H44-3D
-
-
Manually annotated by BRENDA team
strain Y47
-
-
Manually annotated by BRENDA team
strain YSH 11-6B
-
-
Manually annotated by BRENDA team
Sus scrofa Duroc x Yorkshire x Landrace
-
-
-
Manually annotated by BRENDA team
yellow jacket
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
structural and phylogenetic comparisons reveal four main structure types among the five families of glycerol-3-phosphate and glycerol-1-phosphate dehydrogenases, overview
malfunction
-
Drosophila GPDH-1-null mutants cannot fly
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
dihydroxyacetone phosphate + NADH
glycerol-3-phosphate + NAD+
show the reaction diagram
dihydroxyacetone phosphate + NADH
sn-glycerol 3-phosphate + NAD+
show the reaction diagram
dihydroxyacetone phosphate + NADH + H+
sn-glycerol 3-phosphate + NAD+
show the reaction diagram
glycerol-3-phosphate + NAD+
glycerone phosphate + NADH
show the reaction diagram
glycerone phosphate + NADH
glycerol-3-phosphate + NAD+
show the reaction diagram
-
-
-
-
r
glycerone phosphate + NADH
L-glycerol-3-phosphate + NAD+
show the reaction diagram
-
-
-
-
r
glycerone phosphate + NADH
sn-glycerol 3-phosphate + NAD+
show the reaction diagram
-
-
-
-
r
glycolaldehyde + NADH
?
show the reaction diagram
-
-
-
-
?
L-glycerol-3-phosphate + NAD+
glycerone phosphate + NADH
show the reaction diagram
-
-
-
-
r
sn-glycerol 3-phosphate + NAD+
dihydroxyacetone phosphate + NADH + H+
show the reaction diagram
sn-glycerol 3-phosphate + NAD+
glycerone phosphate + NADH + H+
show the reaction diagram
sn-glycerol 3-phosphate + NADP+
dihydroxyacetone phosphate + NADPH + 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
dihydroxyacetone phosphate + NADH
glycerol-3-phosphate + NAD+
show the reaction diagram
dihydroxyacetone phosphate + NADH + H+
sn-glycerol 3-phosphate + NAD+
show the reaction diagram
-
-
-
-
?
sn-glycerol 3-phosphate + NAD+
dihydroxyacetone phosphate + NADH + H+
show the reaction diagram
sn-glycerol 3-phosphate + NAD+
glycerone phosphate + NADH + H+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
-
0.1-0.3 M, chloride salts become strongly inhibitory at higher concentrations, replacing K+ with Na+ or NH4+yields similar results
MgCl2
-
0.1-0.3 M
MgSO4
-
0.1-0.3 M, slight activation at low ionic strength, most inhibitory at higher concentration
NaCl
-
when Dunaliella salina is cultured chronically at various salinities, the accumulation of single cell glycerol increases with increased salinity, Dunaliella salina also can rapidly decrease or increase single cell glycerol contents to adapt to hypoosmotic or hyperosmotic shock
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(-)-epigallocatechin-3-gallate
-
noncompetitive
(NH4)2SO4
2,3-Dimercaptopropanol
-
competitive inhibitor to glycerophosphate
2,4-Dichlorophenoxyacetic acid
-
-
2-amino-2-hydroxymethylpropane-1,3-diol
-
i.e. Tris
2-hydroxy-1,2,3-nonadecanetricarboxylic acid
3-morpholinosydnonimine
-
-
adenosine diphosphate ribose
ADP
-
at physiological concentration, 10 mM 90% inhibition
ADP-ribose
-
allosteric inhibitor
Cd2+
-
50% inhibition at 8.33 mM; 50% inhibition at 8.3 mM
Cu2+
-
50% inhibition at 12.96 mM; 50% inhibition at 13.0 mM
dihydroxyacetone phosphate
FeCl2
-
0.5 mM, about 10% residual activity
fructose-1,6-bisphosphate
-
at physiological concentration, non-competitive
glycerol
-
slight inhibition
glycerol phosphate
-
0.025 mM, strong inhibition of chloroplastic and cytosolic form, endproduct inhibition
gymnemic acid
-
may have some pharmacological activities including antidiabetic activity and lipid lowering effects via interaction with G3PDH
iodoacetamide
-
effective inhibitor
iodoacetate
K+
-
50% inhibition at 40.66 mM; 50% inhibition at 40.7 mM
KCl
-
inactivation
L-glycerol 3-phosphate
Large peptide factor
-
chloroplast enzyme
-
linoleic acid
-
0.003 mM, 40% inhibition of chloroplastic form and 43% inhibition of cytosolic form
malate
-
at high concentration
Melarsen oxide
Mn2+
-
50% inhibition at 36.6 mM
N-ethylmaleimide
NADH-X
-
allosteric inhibitor
Ni2+
-
50% inhibition at 31.66 mM; 50% inhibition at 31.7 mM
Nucleic acids
-
strongly inhibit
-
o-Iodosobenzoic acid
-
0.5 mM, about 10% residual activity
octyl glucose
-
0.003 mM, 41% inhibition of chloroplastic form and 61% inhibition of cytosolic form
p-chloromercuribenzoate
p-mercuribenzoate
-
0.1 mM complete inhibition
palmitic acid
-
0.003 mM, 40% inhibition of chloroplastic form and 43% inhibition of cytosolic form
phenylmethyl sulfonyl fluoride
-
0.5 mM 50% inhibition of cytosolic form, little effect on chloroplastic form
phosphate
phosphatidyl choline
-
0.003 mM, 42% inhibition of chloroplastic form and 43% inhibition of cytosolic form
phosphogluconate
-
cytosolic isozyme
reduced thioredoxin
-
stimulation of chloroplastic form
S-nitroso-N-acetylpenicillamine
-
-
sedoheptulose 1,7-bisphosphate
-
0.5 mM, chloroplastic and cytosolic form inhibited by 50%
selenocysteine
-
0.015 mM, 10% residual activity
selenomethionine
-
0.015 mM, 40% residual activity
Small peptide factor
-
cytosolic enzyme
-
SO42-
-
at high concentration, MgSO4 most inhibitory
Sodium selenite
-
0.015 mM, 10% residual activity
suramin
Thylakoid fraction
-
0.02 mM 97% inhibition
-
Triton X-100
-
0.006 mM, 50% inhibition of chloroplastic form and 52% inhibition of cytosolic form
Zn2+
-
50% inhibition at 5.66 mM; 50% inhibition at 5.7 mM
ZnCl2
-
0.5 mM, about 10% residual activity
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ATP
-
slight activation
bovine serum albumin
-
activity increased
-
Dihydrolipoic acid
-
stimulation of chloroplastic form, 147% of enzyme activity
dipotassium malate
-
slight activation at low ionic strength
dithiothreitol
-
stimulation of chloroplastic form, 150% of enzyme activity
EDTA
-
without EDTA the rates are about 60% of the maximal rate
fructose 2,6-bisphosphate
-
0.025 mM maximum stimulation of 2.3fold of cytosolic form, not chloroplastic form, no stimulation by fructose-1,6-bisphosphate
glycerol
-
competitive activator with respect to dihydroxyacetone phosphate above 30 mM
Glycine buffer
-
0.9 M, slight activation
-
phosphate
-
around 5 mM, activation, chloroplastic form
phosphite dianion
-
allosteric activation. Separate binding of the second substrate piece phosphite dianion strongly activates GPDH for catalysis of the reduction of glycolaldehyde by NADH
phosphogluconate
-
0.005 mM slightly stimulates chloroplastic form
potassium glutamate
-
up to 0.1 M, activation, high activity also at high ionic strength
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.3
alpha-glycerol phosphate
-
-
0.11 - 3.39
alpha-glycerophosphate
0.012 - 0.54
dihydroxyacetone phosphate
0.19 - 1.2
DL-glycerol-3-phosphate
0.59 - 2
glycerol 3-phosphate
1.6
glycerol phosphate
-
-
0.468 - 34
glycerol-3-phosphate
0.137
Glycerone
-
pH 7.6, 25C
0.154 - 137
glycerone phosphate
0.74
L-glycerol 3-phosphate
-
-
2.3 - 3.9
L-glycerol-3-phosphate
0.0044 - 316
NAD+
0.001 - 32.4
NADH
0.0589 - 0.0726
NADPH
469
sn-glycerol 3-phosphate
-
pH 7.6, 25C
0.143
sn-glycerol-3-phosphate
-
measured at pH 9.0
additional information
alpha-glycerophosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
188 - 390
dihydroxyacetone phosphate
19000
glycerone phosphate
Eidolon helvum
-
pH 7.5, 25C
32 - 78
L-glycerol-3-phosphate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02
(-)-epigallocatechin-3-gallate
-
substrate dihydroxyacetone phosphate, pH 7.4, 25C; substrate NADH, pH 7.4, 25C
0.2
gymnemic acid
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.02
(-)-epigallocatechin-3-gallate
Oryctolagus cuniculus
-
pH 7.4, 25C
0.00055 - 0.0011
2-hydroxy-1,2,3-nonadecanetricarboxylic acid
0.0015 - 0.005
Melarsen oxide
Trypanosoma brucei
-
cymelarsen, IC50 0.0015-0.005 mM
0.0002 - 0.00044
suramin
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.1
-
standard deviation 0.2, crude preparation, testes
33
-
isozyme P-V, liver
53.6
-
pH 7.6, 25C
55.6
-
pH 9.0, 25C
70.7
-
standard deviation 5.1, crude preparation, liver
77
-
isozyme I6.5, after affinity chromatography, after preparative isoelectric-focusing: 175 units/mg
80
-
estimated
88.2
-
-
96
-
isozyme P-I, liver
119
-
62 units/mg calculated by using protein concentration determined by dry-weight measurements
120
-
pH 7.5, 25C
123
-
isozyme P-IV, liver
156
-
isozyme GPDH-3
162
-
isozyme I5.9
170
-
standard deviation 45 units/mg
179.8
-
isozyme GPDH-1
180
-
mammary gland
234
-
adipose tissue,147 units/mg refer to protein determination by Kjeldahl nitrogen analysis
253
-
isozyme P-II, liver
257
-
purified enzyme, from liver, pH 8.5, 25C
282
-
purified enzyme, from muscle, pH 8.5, 25C
806
-
isozyme P-III, liver
2396
-
isoform Gut2, pH 6.8
additional information
-
activities with the different isozymes at different temperatures, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.2
-
reduction of dihydroxyacetone phosphate, purified enzyme, decline in activity is more rapid at higher pH values, 7.3-7.5 crude homogenate of enzyme
7.4
-
reduction of dihydroxyacetone-phosphate, isozyme GPDH-3
7.6
-
maximum rate of dihydroxyacetone phosphate reduction
7.7
-
reduction of dihydroxyacetone phosphate; reduction reaction
8.5
-
assay at, oxidation reaction
9.5
-
oxidation of glycerol-phosphate
10
-
optimum pH for NAD+ reduction about pH 10.0
10.2
-
oxidation of glycerol 3-phosphate, in 0.03 glycine buffer
additional information
-
optimum pH values for components P-I, P-II, PIV, P-V range between pH 8.2 and 8.6, liver
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.5
-
highest activity, glycerol-3-phosphate formation
5 - 8.1
-
about half-maximal activity at pH 5.0 and 8.1, reduction of dihydroxyacetone phosphate
5 - 7.5
-
highest activity, glycerol-3-phosphate formation
5.2 - 7.8
-
about half-maximal activity between pH 5.2 and 7.8, reduction of dihydroxyacetone phosphate
5.8 - 7.2
-
reduction of dihydroxyacetone phosphate occurs over a broad range between 5.8 to 7.2
6 - 6.5
-
maximal activities of the three allelic forms are obtained within a pH range of 6.0-6.5 for dihydroxyacetone phosphate reduction
6.5 - 8.2
-
about half-maximal activity at pH 6.5 and 8.2, reduction of dihydroxyacetone phosphate
6.7 - 8.7
-
about 80% of maximal activity at pH 6.7 and 8.7, reduction of dihydroxyacetone phosphate, decrease of activity below and above these pH values
6.8 - 7.2
-
activity falls off rapidly as pH is increased
7 - 8
-
dihydroxyacetone reduction, activity decreases rapidly below pH 7.0 and above pH 8.0, more rapidly at acidic values
7.5 - 8
-
pH-rate profile for the reduction of dihydroxyacetone phosphate shows rather sharp optimum between pH 7.5 and 8.0
7.9 - 8.2
-
maximum activity in pH interval 7.9-8.2
8 - 11
-
about half-maximal activity at pH 8.0 and 11.0, oxidation of L-glycerol-3-phosphate
8 - 10
-
about half-maximal activity at pH 8.0 and 10.0, oxidation of glycerol 3-phosphate, isozymes P-III, liver
8.1 - 9.4
-
about 80% of maximal activity at pH 8.1 and 9.4, oxidation of glycerol-3-phosphate, decrease of activity below and rapidly above these pH values
8.6 - 10
-
about half-maximal activity at pH 8.6 and 10.0, oxidation of glycerol 3-phosphate
10 - 10.5
-
maximal activities of the three allelic forms are obtained within a pH range of 10.0-10.5 for glycerol 3-phosphate oxidation
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
isozyme GPDHf, activity falls off at temperature above 30C
40
-
isozyme GPDHm and GPDHs
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.4
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
; and longissimus dorsi muscle, similar expression pattern that is at a low level at birth and increasing with aging to the highest level at postnatal day 8 in longissinus doris muscle and postnatal day 14 in cerebrum. Weaning decreases the expression level of the GPD1 gene
Manually annotated by BRENDA team
very low expression level
Manually annotated by BRENDA team
-
mGPDH abundance and activity is significantly elevated in prostate cancer cell lines when compared to the normal prostate epithelial cell line PNT1A
Manually annotated by BRENDA team
-
mGPDH abundance and activity is significantly elevated in prostate cancer cell lines when compared to the normal prostate epithelial cell line PNT1A
Manually annotated by BRENDA team
high expression level
Manually annotated by BRENDA team
higher expression level
Manually annotated by BRENDA team
-
adipofibroblast, embryonic cell culture, up to 10fold increase of activity in presence of chicken serum, further increase in presence of embryonic chicken serum
Manually annotated by BRENDA team
adult gonad; adult gonad
Manually annotated by BRENDA team
-
in adult worms on tegument urface, and in metacercaria on tegument and tegumentary cells
Manually annotated by BRENDA team
very low expression level
Manually annotated by BRENDA team
-
mGPDH abundance and activity is significantly elevated in prostate cancer cell lines when compared to the normal prostate epithelial cell line PNT1A
Manually annotated by BRENDA team
and cerebellum, similar expression pattern that is at a low level at birth and increasing with aging to the highest level at postnatal day 8 in longissinus dorsi muscle and postnatal day 14 in cerebrum. Weaning decreases the expression level of the GPD1 gene
Manually annotated by BRENDA team
-
mGPDH abundance and activity is significantly elevated in prostate cancer cell lines when compared to the normal prostate epithelial cell line PNT1A
Manually annotated by BRENDA team
-
mGPDH abundance and activity is significantly elevated in prostate cancer cell lines when compared to the normal prostate epithelial cell line PNT1A
Manually annotated by BRENDA team
-
in the absence of Gpd2, hyperactivation and acrosome reaction are significantly altered, and a few changes in protein tyrosine phosphorylation are also observed during capacitation. GPD2 activity is required for generation of reactive oxygen species in mouse spermatozoa during capacitation, failing which, capacitation is impaired
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
isoform Gdp1p, partly
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000
-
SDS-PAGE
59500
-
gel filtration
62000
-
gel filtration
63500
-
gel filtration
65000
-
gel filtration
67000
-
gel filtration
68000
-
gel filtration
69000
-
66000-72000 Da, gel filtration
72500
-
69000-76000, gel filtration
73000
-
calculated by amino acid analysis
74000
-
electrophoresis in a non-denaturing system
76200
calculated from amino acid sequence
76500
-
74000-79000 Da, SDS-PAGE under non-denaturing conditions
77000
-
gel filtration
77200
calculated from amino acid sequence
78600
-
fluorometric titrations
79500
-
meniscus depletion method
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
-
modification of Cys345 in hydrophobic pocket
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified GlpD is concentrated to 8 mg/ml and crystallized at 4C from a solution containing 0.1 M diammonium hydrogen phosphate, 0.1 M Bicine pH 8.5, and 12% w/v PEG 6000. Structure of the native enzyme, structures of enzyme complexed with substrate analogues phosphoenolpyruvate, glyceric acid 2-phosphate, glyceraldehyde-3-phosphate, and product, dihydroxyacetone phosphate
-
hanging-drop vapor-diffusion method enzyme/NAD+ complex, enzyme/dihydroxyacetonephosphate complex, enzyme/NAD+/dihydroxyacetonephosphate complex
purified recombinant untagged enzyme, hanging drop vapour diffusion method using a reservoir solution consisting of 12% PEG 8000, 0.1 M Tris-HCl, pH 8.5, 0.3 M MgCl2, X-ray diffraction structure determination and analysis at 2.45 A resolution, molecular replacement and modeling
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.8 - 9.9
-
stable at 21C for 15 min; stable for 15 min at 21C
287338
5
-
inactivation after 1 h at 20C
287342
5.7 - 8.5
-
around this range, most stable
287331
6 - 9
-
stable for 16 h at 20C
287342
10
-
inactivation after 1 h at 20C
287342
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21
-
15 min stable; stable at pH 4.8-9.9 for 15 min
30
-
30 min stable, pH 6.6
35
-
most stable at pH 8, activity reduced, GPDHf to a greater extent
61
-
complete inactivation after 5 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol does not stabilize
-
2-mercaptoethanol stabilizes during purification
20% polyethylene glycol stabilizes during purification
ammonium sulfate inactivates
-
ammonium sulfate stabilizes
bovine serum albumin stabilizes
-
charcoal inactivates rabbit enzyme, restorable by thiamic acid
-
dithiothreitol does not stabilize
-
dithiothreitol stabilizes
EDTA stabilizes
even cold dilute solutions in 0.2 M ammonium sulfate at pH 5.8 are stable for weeks
-
freezing inactivates completely, ammonium sulfate prevents inactivation
-
lyophilization inactivates
-
NAD+ stabilizes
-
NADH stabilizes
-
NADH stabilizes preparations of low ionic strength
-
phenylmethane-sulfonyl fluoride stabilizes during purification
-
phosphate buffer stabilizes
-
redistilled water prevents denaturation in absence of salts
-
repeated freezing and thawing results in rapid loss of activity, to some extent restorable at room temperature
-
substrates stabilize dilute preparation
-
the enzyme displays rapid induction as well as decay upon dissappearance of a hormonal stimulus (thyroid hormone), indicating a rather short half-life of the enzyme
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20-0C, stable in 75% ammonium sulfate
-
-20C, 50% glycerol, retains activity for several months
-
-20C, in 0.1 M Tris-HCl, 2 mM 2-mercaptoethanol, pH 7.0, 50% polyethylene glycol 2000, 5 mM NADH or dihydroxyacetone phosphate, unstable
-
-20C, stable as (NH4)2SO4-precipitate
-
-20C, stable for at least 2 months
-
-80C, in 0.1 M Tris-HCl, 2 mM 2-mercaptoethanol, pH 7.0, 50% polyethylene glycol 2000, 5 mM NADH or dihydroxyacetone phosphate, unstable
-
0C, 30 mM Tris-H2SO4, 2.5 mM Na2EDTA, 5 mM dithiothreitol, 0,05% NaN3 (pH 8, 4C), 0.1 mM phenylmethylsulfonylfluoride, 15-17.5% w/v poly(ethylene glycol) 4000, after six days loss of activity insignificant
-
0C, crude extract stable for 6 days in presence of polyethylene glycol
-
0C, t1/2: 50 min
-
2C, crystals stable at least a month in 0.1 M Tris buffer, pH 7.6 with added EDTA, dithiothreitol and ammonium sulfate
-
4C, 1 M Tris-HCl, 2 mM 2-mercaptoethanol, pH 7.0, eluted from Sephadex G-100 column, 90% activity remained after several weeks
-
4C, 20% v/v, glycerol, purified stable
4C, dilute solution, pH 5.8, stable for weeks
-
4C, purified and dilute preparation stable for several days in presence of NADH
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4C, stable
4C, stable for several weeks in 1 M Tris/HCl buffer with 2-mercaptoethanol, pH 7.0
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4C, t1/2: 6 h in absence of EDTA and polyethyleneglycol
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5C, isozyme 1: stable for over a month in 10 mM sodium phosphate buffer, pH 6.5, with added EDTA, dithiothreitol and NADH, isozyme 3: 50% loss of activity under the same conditions
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crystallized, in ammonium sulfate stable for months
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crystals, 2C in 0.1M Tris, 10 mM EDTA, 2 mM dithiothreitol, pH 7.6, (NH4)2SO4 in final concentration of 1.7 M
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 allelic forms of the enzyme; affinity chromatography
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2 isozymes
2 isozymes with distinct isoelectric points in brain
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3 allelic forms distinguishable by electrophoresis
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3 isozymes in adults and 1 in larvae, distinct in isoelectric focusing; affinity chromatography
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3 isozymes, product of the same gene mapped to left arm of chromosome II, isozyme 1: flight muscle, isozyme 3: larval and adult fat body, isozyme 2: heterodimer of 1 and 3; affinity chromatography
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4 isozymes in testes and 5 isozymes in liver, distinct in isoelectric focusing
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6 to 8 isozymes of bumblebee flight muscle enzyme
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affinity chromatography
dye-affinity chromatography
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GSTrap column chromatography; GSTrap column chromatography
ion-exchange chromatography combined with affinity elution, 2 isozymes that differ in charge by analytical PAGE
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native enzyme 50fold and 176fold from liver and muscle, respectively, by anion exchange chromatography, gel filtration, and affinity chromatography
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preparative isoelectric focusing
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recombinant enzyme from Escherichia coli strain BL21 by anion exchange chromatography to over 95% purity
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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sequential affinity chromatography; several isozymes, distinct in isoelectric points
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several isozymes, distinct in isoelectric points
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