Information on EC 3.1.3.75 - phosphoethanolamine/phosphocholine phosphatase

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

EC NUMBER
COMMENTARY hide
3.1.3.75
-
RECOMMENDED NAME
GeneOntology No.
phosphoethanolamine/phosphocholine phosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
O-phosphoethanolamine + H2O = ethanolamine + phosphate
show the reaction diagram
phosphocholine + H2O = choline + phosphate
show the reaction diagram
phosphoethanolamine + H2O = ethanolamine + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
choline biosynthesis I
-
-
Glycerophospholipid metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphoethanolamine phosphohydrolase
Requires active site Mg2+ but also works, to a lesser extent, with Co2+ and Mn2+. The enzyme is highly specific for phosphoethanolamine and phosphocholine.
CAS REGISTRY NUMBER
COMMENTARY hide
52227-92-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
the regulation of choline metabolism and consequently PchP synthesis may reflect an adaptive response of Pseudomonas aeruginosa to environmental conditions. Regulation of pchP gene expression, overview
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
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
show the reaction diagram
-
-
-
-
?
beta-glycerol phosphate + H2O
glycerol + phosphate
show the reaction diagram
-
poor substrate
-
-
?
D-ribose 5-phosphate + H2O
D-ribose + phosphate
show the reaction diagram
-
poor substrate
-
-
?
O-phosphocholine + H2O
choline + phosphate
show the reaction diagram
O-phosphoethanolamine + H2O
ethanolamine + phosphate
show the reaction diagram
p-nitrophenyl phosphate + H2O
nitrophenol + phosphate
show the reaction diagram
p-nitrophenyl phosphate + H2O
p-nitrophenol + phosphate
show the reaction diagram
phospho-L-serine + H2O
L-serine + phosphate
show the reaction diagram
-
-
-
-
?
phosphocholine + H2O
choline + phosphate
show the reaction diagram
phosphoethanolamine + H2O
ethanolamine + phosphate
show the reaction diagram
pyridoxal 5'-phosphate + H2O
pyridoxal + phosphate
show the reaction diagram
-
poor substrate
-
-
?
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
O-phosphocholine + H2O
choline + phosphate
show the reaction diagram
O-phosphoethanolamine + H2O
ethanolamine + phosphate
show the reaction diagram
phosphocholine + H2O
choline + phosphate
show the reaction diagram
phosphoethanolamine + H2O
ethanolamine + phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
divalent metal ions required, highest activity with 1.5 mM Mg2+
Mn2+
-
stimulates to a lesser extend than Mg2+, higher activity with phosphocholine than with phosphoethanolamine in the presence of Co2+ and Mn2+ most probably due to an allosteric effect caused by a difference in the metal-binding properties of each enzyme-substrate complex
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-(([3-(3-oxo-1,2-benzothiazol-2(3H)-yl)phenyl]sulfonyl)amino)benzoic acid
-
-
2-(2,3-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
-
-
2-(2,5-dimethylphenyl)-1,2-benzothiazol-3(2H)-one
-
-
2-(2,5-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
-
-
2-(3-chloro-4-fluorophenyl)-1,2-benzothiazol-3(2H)-one
-
-
2-(3-chlorophenyl)-1,2-benzothiazol-3(2H)-one
-
-
2-(3-methylphenyl)-1,2-benzothiazol-3(2H)-one
-
-
2-(4-fluorophenyl)-1,2-benzothiazol-3(2H)-one
-
-
2-(4-methylphenyl)-1,2-benzothiazol-3(2H)-one
-
-
2-phenyl-1,2-benzoisoselenazol-3(2H)-one
-
noncompetitive inhibitor
2-phenyl-1,2-benzothiazol-3(2H)-one
-
-
2-[2-(morpholin-4-yl)-5-(morpholin-4-ylsulfonyl)phenyl]-1,2-benzothiazol-3(2H)-one
-
-
2-[4-(dimethylamino)phenyl]-1,2-benzothiazol-3(2H)-one
-
-
2-[4-chloro-3-(morpholin-4-ylsulfonyl)phenyl]-1,2-benzothiazol-3(2H)-one
-
-
2-[4-methyl-3-(morpholin-4-ylsulfonyl)phenyl]-1,2-benzothiazol-3(2H)-one
-
-
2-[5-(morpholin-4-ylsulfonyl)-2-(pyrrolidin-1-yl)phenyl]-1,2-benzothiazol-3(2H)-one
-
-
3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzoic acid
-
-
5-fluoro-2-phenyl-1,2-benzothiazol-3(2H)-one
-
-
6-fluoro-2-(4-fluorophenyl)-1,2-benzothiazol-3(2H)-one
-
-
6-fluoro-2-(4-methoxyphenyl)-1,2-benzothiazol-3(2H)-one
-
-
6-fluoro-2-phenyl-1,2-benzothiazol-3(2H)-one
-
-
betaine
choline
ebselen
-
-
ethyl 4-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzoate
-
-
lansoprazole
methyl 3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzoate
-
-
N,N-diethyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzenesulfonamide
-
-
N,N-dimethyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzamide
-
-
N,N-dimethyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzenesulfonamide
-
-
N-benzyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzamide
-
-
phosphocholine
-
substrate inhibition at high concentration
phosphorylcholine
-
SCH202676
Tetramethylammonium chloride
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ascorbic acid
-
increases PHOSPHO1 expression in matrix vesicles after 12 days in culture
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.51 - 2
4-nitrophenyl phosphate
0.003
O-Phosphoethanolamine
-
pH 6.7, 37C, 2 mM Mg2+
1.4 - 22.2
p-nitrophenyl phosphate
0.0114 - 3.6
phosphocholine
1.16
Phosphoethanolamine
50 mM Hepes/NaOH buffer, pH 7.0, 37C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.7 - 87.1
4-nitrophenyl phosphate
2.27
O-Phosphoethanolamine
-
pH 6.7, 37C, 2 mM Mg2+
0.2 - 7900000
p-nitrophenyl phosphate
1.04 - 1.98
phosphocholine
13
Phosphoethanolamine
50 mM Hepes/NaOH buffer, pH 7.0, 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
13.3 - 141
4-nitrophenyl phosphate
24 - 44
p-nitrophenyl phosphate
2.34
phosphocholine
50 mM Hepes/NaOH buffer, pH 7.0, 37C
1.06
Phosphoethanolamine
50 mM Hepes/NaOH buffer, pH 7.0, 37C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.031
ebselen
-
-
0.07128
lansoprazole
-
-
0.00108
SCH202676
-
-
0.00035
Zn2+
-
at pH 7.4
additional information
additional information
inhibition kinetics for alkylammonium compounds trimethylamine, tetramethylammonium chloride, choline, chlorocholine, hexamethonium, and decamethonium, in presence of Zn2+ or Mg2+, overview
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00081
2-(([3-(3-oxo-1,2-benzothiazol-2(3H)-yl)phenyl]sulfonyl)amino)benzoic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.0047
2-(2,3-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.004
2-(2,5-dimethylphenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.003
2-(2,5-dimethylphenyl)-6-fluoro-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0018
2-(3-chloro-4-fluorophenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0052
2-(3-chlorophenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0027
2-(3-methylphenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0049
2-(4-fluorophenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0013
2-(4-methylphenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.00094
2-phenyl-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0012
2-[2-(morpholin-4-yl)-5-(morpholin-4-ylsulfonyl)phenyl]-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.011
2-[4-(dimethylamino)phenyl]-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0011
2-[4-chloro-3-(morpholin-4-ylsulfonyl)phenyl]-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0018
2-[4-methyl-3-(morpholin-4-ylsulfonyl)phenyl]-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0075
2-[5-(morpholin-4-ylsulfonyl)-2-(pyrrolidin-1-yl)phenyl]-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0011
3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzoic acid
Homo sapiens
-
pH and temperature not specified in the publication
0.00094
5-fluoro-2-phenyl-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.01
6-fluoro-2-(4-fluorophenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.0067
6-fluoro-2-(4-methoxyphenyl)-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.00074
6-fluoro-2-phenyl-1,2-benzothiazol-3(2H)-one
Homo sapiens
-
pH and temperature not specified in the publication
0.00281
ebselen
Homo sapiens
-
-
0.0012
ethyl 4-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzoate
Homo sapiens
-
pH and temperature not specified in the publication
0.00471
lansoprazole
Homo sapiens
-
-
0.00082
methyl 3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzoate
Homo sapiens
-
pH and temperature not specified in the publication
0.00056
N,N-diethyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzenesulfonamide
Homo sapiens
-
pH and temperature not specified in the publication
0.00014
N,N-dimethyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzamide
Homo sapiens
-
pH and temperature not specified in the publication
0.0005
N,N-dimethyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzenesulfonamide
Homo sapiens
-
pH and temperature not specified in the publication
0.0023
N-benzyl-3-(3-oxo-1,2-benzothiazol-2(3H)-yl)benzamide
Homo sapiens
-
pH and temperature not specified in the publication
0.00197
SCH202676
Homo sapiens
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0176
-
pH 7.2, 37C, hydrolysis of pyridoxal 5-phosphate
0.0396
-
pH 7.2, 37C, hydrolysis of beta-glycerol phosphate
0.0645
-
pH 7.2, 37C, hydrolysis of p-nitrophenyl phosphate
0.0748
-
pH 7.2, 37C, hydrolysis of ribose 5-phosphate
0.092
purified recombinant mutant D43N enzyme fragment M19-C267, substrate O-phosphoethanolamine
0.28
purified recombinant mutant D123N enzyme fragment M19-C267, substrate O-phosphoethanolamine
0.48
substrate phosphocholine, 50 mM Hepes/NaOH buffer, pH 7.0, 37C
1.3
-
mutant D262E, at pH 5, with p-nitrophenyl phosphate as substrate
1.5
-
mutant D267E, at pH 5, with p-nitrophenyl phosphate as substrate
2.4
-
mutant D267E, at pH 5, with phosphocholine as substrate
2.5
-
mutant D267E, at pH 7.4, with phosphocholine as substrate
2.98
-
pH 7.2, 37C, hydrolysis of phosphocholine
3
-
mutant T35S, at pH 5, with phosphocholine as substrate
3.6
-
mutant T35S, at pH 7.4, with phosphocholine as substrate
3.67
substrate phosphoethanolamine, 50 mM Hepes/NaOH buffer, pH 7.0, 37C
4.6
-
pH 7.2, 37C, hydrolysis of phosphoethanolamine
4.96
purified recombinant wild-type enzyme fragment M19-C267, substrate O-phosphoethanolamine
5.9
-
mutant T35S, at pH 5, with p-nitrophenyl phosphate as substrate
6.5
-
mutant D262E, at pH 5, with phosphocholine as substrate
7
-
mutant S166T, at pH 5, with p-nitrophenyl phosphate as substrate
10.1
-
purified recombinant mutant Y82A/Y83A/Y84A, pH 5.0, 37C, substrate 4-nitrophenyl phosphate
11.1
-
mutant D262E, at pH 7.4, with phosphocholine as substrate
20.3
-
mutant S166T, at pH 5, with phosphocholine as substrate
26.2
-
mutant S166T, at pH 7.4, with phosphocholine as substrate
38.8
-
mutant G261A, at pH 5, with phosphocholine as substrate
54.5
-
substrate phosphocholine, with 2 mM Mg2+
56.9
-
mutant K242R, at pH 5, with phosphocholine as substrate
59
-
purified recombinant mutant E42A, pH 5.0, 37C, substrate 4-nitrophenyl phosphate
63.4
-
mutant G261A, at pH 5, with p-nitrophenyl phosphate as substrate
64
-
mutant D265E, at pH 5, with p-nitrophenyl phosphate as substrate
64.8
-
mutant G261A, at pH 7.4, with phosphocholine as substrate
70
-
mutant D265E, at pH 7.4, with phosphocholine as substrate
71
-
mutant D265E, at pH 5, with phosphocholine as substrate
74.5
-
wild-type with motif 31DMDNT35, at pH 5, with p-nitrophenyl phosphate as substrate
74.6
-
mutant K242R, at pH 7.4, with phosphocholine as substrate
75
-
wild type enzyme, at pH 5.0 and 37C, in the presence of 2 mM Mg2+
86.9
-
wild-type with motif 31DMDNT35, at pH 7.4, with phosphocholine as substrate
88.4
-
wild-type with motif 31DMDNT35, at pH 5, with phosphocholine as substrate
100
-
wild-type with motif 166S, motif 242K or motif 161GDTPDSD267, at pH 5, with p-nitrophenyl phosphate as substrate and at pH 5 and pH 7.4 with phosphocholine as substrate
106.7
-
mutant K242R, at pH 5, with p-nitrophenyl phosphate as substrate
110
-
substrate p-nitrophenyl phosphate
114
-
wild type enzyme, at pH 5.0 and 37C, in the presence of 0.3 mM Cu2+
121
-
purified recombinant mutant E43A, pH 5.0, 37C, substrate 4-nitrophenyl phosphate
132.7
-
purified recombinant wild-type enzyme, pH 5.0, 37C, substrate 4-nitrophenyl phosphate
142
-
wild type enzyme, at pH 5.0 and 37C, in the presence of 0.5 mM Zn2+
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
maximum of kcat-value and minimum for KM-value
7.4
-
assay at, substrates O-phosphocholine and phospho-L-serine
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
optimal range, substrate phosphocholine
6 - 7.5
-
above 70% of maximum activity between pH 6 and 7.2, activity with phosphocholine declines significantly above pH 7.2, activity with phosphoethanolamine declines above pH 7.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
theoretical value
7.1
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
low expression, 3.38fold higher than in liver
Manually annotated by BRENDA team
-
mineralization by PHOSPHO1 in medullary bone
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
from 17 days old embryonic chickens, PHOSPHO1 gene expression
Manually annotated by BRENDA team
from 3 weeks old male chickens, from tibia, growth plate cartilage, expression pattern, PHOSPHO1 is specifically expressed at sites of mineralization in bone and cartilage
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
from growth plates of proximal tibiotarsi, 3X11A expression at high levels in differentiating growth plate chondrocytes, 3X11A expression is upregulated 5fold during chondrocyte terminal differentiation and its expression is approximately 100fold higher in hypertrophic chondrocytes than in non-chondrogenic tissues
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
-
PHOSPHO1 expression occurs prior to E6.5 and is initially restricted to the bone collar within the mid-shaft of the diaphysis of long bones. By E11.5, expression is observed over the entire length of the diaphysis
Manually annotated by BRENDA team
-
low expression, 1.2fold higher than in liver
Manually annotated by BRENDA team
-
low expression, 5.67fold higher than in liver
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
expression of PHOSPHO1
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28600
x * 33000, recombinant PHOSPHO1, SDS-PAGE, x * 28600, x * 30400, alternative transcripts for PHOSPHO1 in growth cartilage, Western blot analysis
29000
-
PHOSPHO1 splice variant, Western blot analysis
29700
x * 29700, sequence calculation
30400
x * 33000, recombinant PHOSPHO1, SDS-PAGE, x * 28600, x * 30400, alternative transcripts for PHOSPHO1 in growth cartilage, Western blot analysis
30442
x * 30442, sequence calculation
31570
calculated from amino acid sequence
33000
x * 33000, recombinant PHOSPHO1, SDS-PAGE, x * 28600, x * 30400, alternative transcripts for PHOSPHO1 in growth cartilage, Western blot analysis
37000
-
2 * 37000, about, recombinant His6-tagged enzyme, SDS-PAGE
37089
-
2 * 37089, MS, multiangle light scattering
60000
-
about, recombinant His6-tagged enzyme, gel filtration
64000
native gel elelctrophoresis, gel filtration
74600
-
multiangle light scattering
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
2 * 37089, MS, multiangle light scattering
monomer
1 * 32000, monomer-dimer equilibrium, both the monomer and the dimer are active
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
D31 is phosphorylated during phosphoester hydrolysis
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop method
-
purified recombinant enzyme, sitting drop vapour diffusion method, 250 nl of 10 mg/ml protein in 200 mM NaCl, 10 mM Tris-HCl, pH 8.0, are mixed with 250 nl well solution and are equilibrated against 0.5 ml well solution containing 0.1 M Bis-Tris, pH 5.5, 0.2 M MgCl2, 25% PEG 3350, producing clustered plate-like microcrystals, or by microbatch method mixing 0.001 ml of protein solution containing 10 mg/ml in 200 mM NaCl, 20 mM Bis-Tris, pH 5.5, with 0.002 ml precipitant solution, cryoprotection by a quick soak in 0.1 M Bis-Tris pH 5.5, 0.2 M MgCl2, 25% PEG 3350, 15% glycerol, X-ray diffraction structure determination and analysis at 2.7 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
apparent Tm value: 65.8C (5 mM Mg2+)
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10C, 10 mM Tris-HCl, pH 8, 30% glycerol
-
-20C, 10 mM Tris-HCl, pH 8, containing 30% glycerol, more than 1 year, no loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from solubilized inclusion bodies, ion-exchange chromatography (Q-Sepharose), dialysis, gel filtration
-
immobilized metal ion affinity chromatography (Ni2+)
-
Ni2+-Sepharose activated column
recombinant enzyme from Escherichia coli by chitin affinity chromatography
-
recombinant His-tagged wild-type and mutant fragments M19-C267 from Escherichia coli by nickel affinity chromatography to homogeneity
recombinant N-terminally His6-tagged PchP from Escherichia coli by nickel affinity chromatography
-
recombinant PchP from Escherichia coli, as an N-terminal fusion to intein or a histidine tag, in the folded state
recombinant PHOSPHO1
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21
expressed in Escherichia coli BL21(DE3)
-
expressed in Escherichia coli ER2566 cells
-
expression in Escherichia coli TOP10
-
expression of His-tagged wild-type and mutant fragment comprising residues M19-C267 in Escherichia coli
expression of wild-type and mutant enzymes in Escherichia coli strain BL21
-
from growth plate chondrocytes, expression in Escherichia coli TOP10
gene PA5292, expression in Escherichia coli, expression in and complementation of a Pseudomonas aeruginosa mutant strain JUF8-00
-
gene pchP, expression of His-tagged enzyme in Escherichia coli strain BL21
gene pchP, phylogenetic tree, regulation of pchP gene expression is under the control of GbdR and NtrC. PchP expression in Escherichia coli as an N-terminal fusion to intein or a histidine tag
gene pchP327, recombinant expression with a N-terminal fusion to an MGSSHHHHHHSSGLVPRGSH tag in Escherichia coli BL21, expression of N-terminal His6-tagged PchP in Escherichia coli
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His-tagged version expressed in Escherichia coli BL21 Codonplus
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into vector pBAD TOPO TA, expressed in Escherichia coli
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overexpression of the wild-type and mutant enzymes in Escherichia coli ER2566, transformed with pTY-pchP327, pTYpchP349, or the mutated expression plasmids
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
dimethylglycine is likely the metabolite directly involved in the induction of PchP. Induction of pchP transcription by glycine betaine, a product of choline oxidation, via betaine aldehyde, mediated by GbdR, an AraC family transcription factor
the regulation of pchP gene expression is under the control of GbdR and NtrC
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D123N
site-directed mutagenesis, the mutant enzyme fragment shows reduced activity compared to the wild-type enzyme fragment
D203S
site-directed mutagenesis, inactive mutant enzyme fragment
D32N
site-directed mutagenesis, inactive mutant enzyme fragment
D43N
site-directed mutagenesis, the mutant enzyme fragment shows reduced activity compared to the wild-type enzyme fragment
D43N/D123N
site-directed mutagenesis, inactive mutant enzyme fragment
C87A
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involved in a non-essential intramolecular disulfide bond
C94A
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involved in a non-essential intramolecular disulfide bond
D265E
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enzyme activity reduced to 30-35% of wild-type activity
D31E
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protein expression, but no enzyme activity
D33E
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protein expression, but no enzyme activity
E42A
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site-directed mutagenesis
E43A
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site-directed mutagenesis
G261A
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reduced activity as compared to wild-type
K242Q
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is inactive
K242R
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functional enzyme
Y82A/Y83A/Y84A
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site-directed mutagenesis
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
dialysis, buffer with 5 mM Mg2+
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine