Information on EC 3.1.3.1 - alkaline phosphatase and Organism(s) Homo sapiens and UniProt Accession P05186

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Homo sapiens
UNIPROT: P05186
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The expected taxonomic range for this enzyme is: Archaea, Eukaryota, Bacteria


The taxonomic range for the selected organisms is: Homo sapiens

EC NUMBER
COMMENTARY hide
3.1.3.1
-
RECOMMENDED NAME
GeneOntology No.
alkaline phosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a phosphate monoester + H2O = an alcohol + phosphate
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
diethylphosphate degradation
-
-
NAD phosphorylation and dephosphorylation
-
-
NAD salvage pathway III
-
-
NAD/NADH phosphorylation and dephosphorylation
-
-
sulfopterin metabolism
-
-
Aminobenzoate degradation
-
-
Folate biosynthesis
-
-
Metabolic pathways
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphate-monoester phosphohydrolase (alkaline optimum)
Wide specificity. Also catalyses transphosphorylations. The human placental enzyme is a zinc protein. Some enzymes hydrolyse diphosphate (cf. EC 3.6.1.1 inorganic diphosphatase)
CAS REGISTRY NUMBER
COMMENTARY hide
9001-78-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
-
in neonatal blood plasma, soluble 5'-nucleotidase and alkaline phosphatase mediate conversion of AMP to adenosine, whereas soluble adenosine deaminase catabolizes adenosine to inosine. Newborn blood plasma demonstrates substantially higher adenosine-generating 5'-nucleotidase and alkaline phosphatase activity and lower metabolizing adenosine deaminase activity than adult plasma. Abundant alkaline phosphatase expressed on the surface of circulating neonatal neutrophils is the dominant AMPase on these cells
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-AMP + H2O
adenosine + phosphate
show the reaction diagram
-
-
-
-
?
2-chloro-5-(4-methoxyspiro (1,2-dioxetane-3,2'-(5'-chloro)tricyclo[3.3.1.13,7]decan)-4-yl)phenyl phosphate + H2O
2-chloro-5-(4-methoxyspiro (1,2-dioxetane-3,2'-(5'-chloro)tricyclo[3.3.1.13,7]decan)-4-yl)phenol + phosphate
show the reaction diagram
-
CDP-star
-
-
?
2-naphthyl phosphate + H2O
2-naphthol + phosphate
show the reaction diagram
-
-
-
-
?
2-phosphoglycerate + H2O
glycerol + phosphate
show the reaction diagram
-
-
-
-
?
3'-AMP + H2O
adenosine + phosphate
show the reaction diagram
-
-
-
-
?
3-phosphoglycerate + H2O
glycerol + phosphate
show the reaction diagram
-
-
-
-
?
4-methylumbelliferyl phosphate + H2O
4-methylumbelliferone + phosphate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
show the reaction diagram
5'-AMP + H2O
adenosine + phosphate
show the reaction diagram
-
-
-
-
?
5'-dTMP + H2O
?
show the reaction diagram
-
-
-
-
?
5'-UMP + H2O
uridine + phosphate
show the reaction diagram
-
-
-
-
?
ADP + H2O
?
show the reaction diagram
-
-
-
-
?
AMP + H2O
adenosine + phosphate
show the reaction diagram
-
-
-
-
?
ATP + H2O
?
show the reaction diagram
-
-
-
-
?
diphosphate + H2O
2 phosphate
show the reaction diagram
glucose 1-phosphate + H2O
glucose + phosphate
show the reaction diagram
-
-
-
-
?
glycogen synthase + H2O
?
show the reaction diagram
-
-
-
-
?
histone + H2O
?
show the reaction diagram
-
32P-labelled
-
-
?
indoxyl phosphate + nitroblue tetrazolium chloride
nitrioblue diformazan + phosphate
show the reaction diagram
-
-
-
?
p-nitrophenyl phosphate + H2O
p-nitrophenol + phosphate
show the reaction diagram
p-toluidinium 5-bromo-4-chloro-3-indolyl phosphate + H2O
?
show the reaction diagram
-
-
-
-
?
phenyl phosphate + H2O
phenol + phosphate
show the reaction diagram
-
-
-
-
?
phospho-DL-Thr + H2O
phosphate + Thr
show the reaction diagram
-
-
-
-
?
phospho-DL-Tyr + H2O
phosphate + Tyr
show the reaction diagram
-
-
-
-
?
phospho-Ser histone + H2O
?
show the reaction diagram
-
-
-
-
?
phospho-Ser-casein + H2O
?
show the reaction diagram
-
-
-
-
?
phosphoethanolamine + H2O
phosphate + ethanolamine
show the reaction diagram
-
-
-
-
?
phosphorylase kinase + H2O
?
show the reaction diagram
-
-
-
-
?
phosphorylated acetyl-CoA carboxylase + H2O
acetyl-CoA carboxylase + phosphate
show the reaction diagram
-
-
-
-
?
phosphoserine + H2O
phosphate + Ser
show the reaction diagram
-
-
-
-
?
phosvitin + H2O
?
show the reaction diagram
-
-
-
-
?
protamine + H2O
?
show the reaction diagram
-
-
-
-
?
pyridoxal 5'-phosphate + H2O
?
show the reaction diagram
-
-
-
-
?
pyridoxal 5'-phosphate + H2O
pyridoxal + phosphate
show the reaction diagram
-
-
-
?
thiamin diphosphate + H2O
thiamin phosphate + phosphate
show the reaction diagram
-
-
-
-
?
triphosphate + H2O
diphosphate + phosphate
show the reaction diagram
-
-
-
-
?
UDP + H2O
?
show the reaction diagram
-
-
-
-
?
UMP + H2O
uridine + phosphate
show the reaction diagram
-
-
-
-
?
UTP + H2O
?
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
diphosphate + H2O
2 phosphate
show the reaction diagram
-
-
-
?
pyridoxal 5'-phosphate + H2O
pyridoxal + phosphate
show the reaction diagram
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
-
stimulates glycogen synthase phosphatase activity
Zn
-
enzyme contains Zn; enzyme from placenta conains 2-3 Zn per dimer
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,4-dimethoxy-2-methylbenzene
-
-
1-(3,4-dihydroxyphenyl)-2-(1H-imidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(2-ethyl-1H-imidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(2-methyl-1H-benzimidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(2-methyl-1H-imidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(2-phenyl-1H-imidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(4-methyl-1H-imidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(4-methyl-1H-pyrazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(4H-1,2,4-triazol-4-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(5,6-dimethyl-1H-benzimidazol-1-yl)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-(propan-2-ylamino)ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-[(4,6-dimethylpyrimidin-2-yl)sulfanyl]ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-[(4-methyl-5-phenyl-4H-1,2,4-triazol-3-yl)sulfanyl]ethanone
-
-
1-(3,4-dihydroxyphenyl)-2-[[1-(4-methoxyphenyl)-1H-tetrazol-5-yl]sulfanyl]ethanone
-
-
1-bromotetramisole
-
-
1-chloro-4-ethoxy-2-methylbenzene
-
-
1-chloro-4-methoxy-2-methylbenzene
-
-
1-fluoro-4-methoxybenzene
-
-
2,5-dimethoxy-N-(pyridin-3-yl)benzenesulfonamide
-
-
2,5-dimethoxy-N-(quinolin-3-yl)benzenesulfonamide
-
-
2-(1H-benzimidazol-1-yl)-1-(3,4-dihydroxyphenyl)ethanone
-
-
2-(1H-benzimidazol-2-ylamino)-1-(3,4-dihydroxyphenyl)ethanone
-
-
2-(4-bromo-1H-imidazol-1-yl)-1-(3,4-dihydroxyphenyl)ethanone
-
-
2-(4-bromo-2-methyl-1H-imidazol-1-yl)-1-(3,4-dihydroxyphenyl)ethanone
-
-
2-chloro-1,4-dimethoxybenzene
-
-
2-ethoxy-5-methyl-N-(pyridin-3-yl)benzenesulfonamide
-
-
2-methoxy-4-nitro-N-(pyridin-3-yl)benzenesulfonamide
-
-
2-methoxy-4-nitro-N-(quinolin-3-yl)benzenesulfonamide
-
-
2-methoxy-5-methyl-N-(pyridin-3-yl)benzenesulfonamide
-
-
2-methoxy-5-methyl-N-(quinolin-3-yl)benzenesulfonamide
-
-
2-[[2-(3,4-dihydroxyphenyl)-2-oxoethyl]sulfanyl]-4-(methoxymethyl)-6-methylpyridine-3-carbonitrile
-
-
3-[2-(3,4-dihydroxyphenyl)-2-oxoethyl]-6,7-dimethoxy-2-benzofuran-1(3H)-one
-
-
4-bromo-2,5-dimethoxy-N-(pyridin-3-yl)benzenesulfonamide
-
-
4-methoxy-1,2-dimethylbenzene
-
-
4-[[(4,6-dimethylpyrimidin-2-yl)amino]methyl]benzene-1,2-diol
-
-
5-bromo-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide
-
-
5-chloro-2-ethoxy-N-(pyridin-3-yl)benzenesulfonamide
-
-
-
arsenate
-
-
Be2+
-
-
Ca2+
-
10 mM, glycogen synthase phosphatase activity
Cys
-
uncompetitive inhibition of phosphatase actvity, mixed-competitive inhibition of inorganic pyrophosphatase activity. Inorganic pyrophosphatase activity is inhibited more than phosphatase activity. Ca2+ and Mg2+ ion concentrations may regulate this inhibition
diethyl-p-nitrophenyl phosphate
-
-
F-
-
50 mM KF, 13% inhibition of p-nitrophenyl phosphate hydrolysis, 50% inhibition of dephosphorylation of P-labelled histones
heparin
-
histone phosphatase activity
histidine
-
-
L-amino acids
specific uncompetitive inhibition, molecular mechanism involves Arg166 and Glu429
L-homoarginine
L-leucine
-
allozyme D is highly sensitive and shows uncompetitive inhibition, allozymes S and F are less sensitive, allozymes SD and FD, mixed type, respond in an intermediate fashion to inhibition
L-Phe-Gly-Gly
Leu
-
L-Leu, uncompetitive, D-Leu with greatly decreased efficiencies
leucinamide
-
-
leucinol
-
-
levamisole
Mg2+
-
-
NaCN
-
-
NADH
-
-
NEM
-
very slowly
p-hydroxymercuribenzoate
-
reactivation by DTT and 2-mercaptoethanol
Phenanthroline
-
-
phenyl phosphonate
-
competitive
phenylalanine
-
-
phenylphosphonate
-
10 mM, 47% inhibition of p-nitrophenyl phosphate hydrolysis, 80% inhibition of dephosphorylation of P-labelled histones
phosphate
-
-
polylysine
-
histone phosphatase activity
protamine
-
histone phosphatase activity
Trimetaphosphate
-
-
vanadate
-
-
Zn2+
-
above 0.02 mM
additional information
-
no inhibition at 10 mM by L-phenylalanine at 10 mM for all three isozymes
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
albumin
-
3fold stimulation of hydrophilic enzyme, slight stimulation of amphiphilic enzyme
-
phosphatidylcholine
-
1fold stimulation of hydrophilic enzyme form
phosphatidylethanolamine
-
1fold stimulation of hydrophilic enzyme form
phosphatidylinositol
-
1.5fold stimulation of hydrophilic enzyme form
phosphatidylserine
-
1fold stimulation of amphiphilic enzyme form, 3fold stimulation of hydrophilic enzyme form
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5
2'-AMP
-
-
0.01 - 0.177
2-chloro-5-(4-methoxyspiro (1,2-dioxetane-3,2'-(5'-chloro)tricyclo[3.3.1.13,7]decan)-4-yl)phenyl phosphate
0.026 - 0.078
2-naphthyl phosphate
2.3
3'-AMP
-
-
0.0156 - 49.7
4-nitrophenyl phosphate
3
5'-AMP
-
-
4.5
5'-dTMP
-
-
4.4
ADP
-
-
5.7
ATP
-
-
1.4 - 3
beta-Glycerophosphate
0.12 - 4
diphosphate
4.8
glucose 1-phosphate
-
-
0.02 - 5.02
p-nitrophenyl phosphate
0.39 - 0.9
phospho-DL-Thr
0.4 - 0.72
phospho-DL-Tyr
0.38 - 0.68
phospho-L-Ser
2.9
Phosphoethanolamine
-
-
0.12
pyridoxal 5'-phosphate
-
allozyme D, pH 7.5, 37C, in presence of Mg2+ and Zn2+
0.2 - 0.37
pyridoxal phosphate
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
34 - 971
4-nitrophenyl phosphate
244
diphosphate
Homo sapiens
-
allozyme D, pH 7.5, 37C, in presence of Mg2+ and Zn2+
92
pyridoxal 5'-phosphate
Homo sapiens
-
allozyme D, pH 7.5, 37C, in presence of Mg2+ and Zn2+
additional information
additional information
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00149
1,4-dimethoxy-2-methylbenzene
Homo sapiens
-
-
0.0012 - 0.0302
1-(3,4-dihydroxyphenyl)-2-(1H-imidazol-1-yl)ethanone
0.0042
1-(3,4-dihydroxyphenyl)-2-(2-ethyl-1H-imidazol-1-yl)ethanone
Homo sapiens
-
-
0.0012 - 0.0263
1-(3,4-dihydroxyphenyl)-2-(2-methyl-1H-benzimidazol-1-yl)ethanone
0.0033 - 0.0783
1-(3,4-dihydroxyphenyl)-2-(2-methyl-1H-imidazol-1-yl)ethanone
0.0172
1-(3,4-dihydroxyphenyl)-2-(2-phenyl-1H-imidazol-1-yl)ethanone
Homo sapiens
-
-
0.0058 - 0.0411
1-(3,4-dihydroxyphenyl)-2-(4-methyl-1H-imidazol-1-yl)ethanone
0.0027 - 0.0533
1-(3,4-dihydroxyphenyl)-2-(4-methyl-1H-pyrazol-1-yl)ethanone
0.0134
1-(3,4-dihydroxyphenyl)-2-(4H-1,2,4-triazol-4-yl)ethanone
Homo sapiens
-
-
0.0058
1-(3,4-dihydroxyphenyl)-2-(5,6-dimethyl-1H-benzimidazol-1-yl)ethanone
Homo sapiens
-
-
0.0393
1-(3,4-dihydroxyphenyl)-2-(propan-2-ylamino)ethanone
Homo sapiens
-
-
0.0006 - 0.0495
1-(3,4-dihydroxyphenyl)-2-[(4,6-dimethylpyrimidin-2-yl)sulfanyl]ethanone
0.0067
1-(3,4-dihydroxyphenyl)-2-[(4-methyl-5-phenyl-4H-1,2,4-triazol-3-yl)sulfanyl]ethanone
Homo sapiens
-
-
0.002 - 0.008
1-(3,4-dihydroxyphenyl)-2-[[1-(4-methoxyphenyl)-1H-tetrazol-5-yl]sulfanyl]ethanone
0.00069
1-chloro-4-ethoxy-2-methylbenzene
Homo sapiens
-
-
0.00051
1-chloro-4-methoxy-2-methylbenzene
Homo sapiens
-
-
0.00129
1-fluoro-4-methoxybenzene
Homo sapiens
-
-
0.00106
2,5-dimethoxy-N-(pyridin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.00019
2,5-dimethoxy-N-(quinolin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.0008 - 0.018
2-(1H-benzimidazol-1-yl)-1-(3,4-dihydroxyphenyl)ethanone
0.0062 - 0.0938
2-(1H-benzimidazol-2-ylamino)-1-(3,4-dihydroxyphenyl)ethanone
0.0158 - 0.0475
2-(4-bromo-1H-imidazol-1-yl)-1-(3,4-dihydroxyphenyl)ethanone
0.0049 - 0.0087
2-(4-bromo-2-methyl-1H-imidazol-1-yl)-1-(3,4-dihydroxyphenyl)ethanone
0.00113
2-chloro-1,4-dimethoxybenzene
0.00316
2-methoxy-4-nitro-N-(pyridin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.00074
2-methoxy-4-nitro-N-(quinolin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.00185
2-methoxy-5-methyl-N-(pyridin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.00065
2-methoxy-5-methyl-N-(quinolin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.0022 - 0.0048
2-[[2-(3,4-dihydroxyphenyl)-2-oxoethyl]sulfanyl]-4-(methoxymethyl)-6-methylpyridine-3-carbonitrile
0.003 - 0.0292
3-[2-(3,4-dihydroxyphenyl)-2-oxoethyl]-6,7-dimethoxy-2-benzofuran-1(3H)-one
0.00155
4-bromo-2,5-dimethoxy-N-(pyridin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.00168
4-methoxy-1,2-dimethylbenzene
Homo sapiens
-
-
0.0512
4-[[(4,6-dimethylpyrimidin-2-yl)amino]methyl]benzene-1,2-diol
Homo sapiens
-
-
0.00012
5-bromo-2-methoxy-N-(quinolin-3-yl)benzenesulfonamide
Homo sapiens
-
-
0.00054
5-chloro-2-ethoxy-N-(pyridin-3-yl)benzenesulfonamide
Homo sapiens
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
629
-
purified recombinant enzyme expressed in Sf-9 cells
738
-
purified native placental enzyme
850
-
purified recombinant enzyme expressed in Tn-5B1-4 cells
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9 - 9.5
-
hydrolysis of organic diphosphates
9.2
-
hydrolysis of 2 mM p-nitrophenyl phosphate, borate buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
-
pH 6.0: about 30% of maximal activity, pH 9.0: about 15% of maximal activity
9.5 - 11.5
-
pH 9.5: about 40% of maximal activity, pH 11.5: about 45% of maximal activity, hydrolysis of beta-glycerophosphate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
using a monoclonal antibody against bone specific alkaline phosphatase (BAP), its expression is detected in approximately 50% of circulating B cells but not T cells. Following stimulations with Pokeweed Mitogen (PWM) and Staphylococcus aureus Cowan I (SAC), BAP expression on human B cells is increased
Manually annotated by BRENDA team
-
78 160 cells from brain produce the intestinal isoenzyme
Manually annotated by BRENDA team
-
neuronal and endothelial cells exclusively express the bone TNAP transcript
Manually annotated by BRENDA team
-
HeLa S3 cells from cervix produce the term-placental isoenzyme
Manually annotated by BRENDA team
-
HT-29 cells from colon produce the intestinal isoenzyme
Manually annotated by BRENDA team
-
neutrophilic from peripheral blood
Manually annotated by BRENDA team
-
LoVo cells from colon produce the term-placental and the intestinal isoenzyme
Manually annotated by BRENDA team
-
non-carcinoma tissue. Liver type alkaline phosphatesse AP is detected in the heavy and light surfactant aggregates from both non-carcinoma and squamous carcinoma tissues, but not in heavy surfactant aggregates from adenocarcinoma tissues. A larger amount of bone type alkaline phosphatase is found in light surfactant aggregate fractions from squamous cell carcinomas than those from adenocarcinoma tissues or noncarcinoma tissues from patients with either type of cancer
Manually annotated by BRENDA team
-
neuronal and endothelial cells exclusively express the bone TNAP transcript
Manually annotated by BRENDA team
-
activity increases during adipogenesis
Manually annotated by BRENDA team
-
non-carcinoma tissue. Liver type alkaline phosphatesse AP is detected in the heavy and light surfactant aggregates from both non-carcinoma and squamous carcinoma tissues, but not in heavy surfactant aggregates from adenocarcinoma tissues. A larger amount of bone type alkaline phosphatase is found in light surfactant aggregate fractions from squamous cell carcinomas than those from adenocarcinoma tissues or noncarcinoma tissues from patients with either type of cancer
Manually annotated by BRENDA team
-
SW-620 cells from colon produce the term-placental and the liver/bone/kidney isoenzyme
Manually annotated by BRENDA team
-
T24 cells from bladder produce the term-placental isoenzyme
Manually annotated by BRENDA team
-
enzyme activity of alkaline phosphatase is not observed in chondrocytes of the subperichondral and central cartilage layer of thyroid cartilage prior to cartilage mineralization. In the thyroid cartilage of a male foetus from 23rd gestational week, as well as from a 9-year-old girl and an 11-year-old boy, the enzymohistochemical reaction is negative. Beginning approximately at the middle of the second decade, the reaction product of the enzyme alkaline phosphatase is detected in distinct chondrocytes in the neighbourhood of mineralized cartilage areas or within the mineralized cartilage. In a comparison of aged-matched thyroid cartilages, male thyroid cartilage often contains more chondrocytes with a positive enzymohistochemical reaction for alkaline phosphatase than samples from females
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
-
the enzyme from liver is associated with the particulate fractions such as nuclei, mitochondria and lysosomes
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
-
-
-
-
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65000
-
about, recombinant enzyme, gel filtration
66000
-
immunoblotting, immature form bearing high mannose-type N-linked oligosaccharides
66200
-
native placental enzyme, gel filtration
80000
-
immunoblotting, mature form bearing complex-type oligosaccharides
126000
-
isozyme B/I, native PAGE
136000
-
isozyme B1, native PAGE
141000
-
isozyme B2, native PAGE
146000
-
gel filtreation
158000
-
gel filtration
200000
-
gel filtration
211000
-
hydrophilic liver enzyme, density gradient gel electrophoresis
214000
-
hydrophilic enzyme form, gradient gel electrophoresis
220000
-
hydrophilic enzyme from osteosarcoma cells, density gradient gel electrophoresis
227000
-
hydrophilic enzyme from lymphoma cells L248, density gradient gel electrophoresis
255000
-
hydrophilic enzyme form, gel filtration
340000
-
gel filtration
433000
-
amphiphilic liver enzyme, density gradient gel electrophoresis
483000
-
amphiphilic enzyme from osteosarcoma cells, density gradient gel electrophoresis
494000
-
amphiphilic enzyme from lymphoma cells L248, density gradient gel electrophoresis
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
sucrose density gradient centrifugation, wild-type and V406A form a homodimer
monomer
-
1 * 64000, recombinant enzyme, SDS-PAGE, 1 * 66200, native enzyme, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
mature form of 80 ka bears complex-type oligosaccharides. The 80 kDa form is sensitive to neuraminidase, but is resistant to treatment with Endo H
glycoprotein
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
10-16 mg/ml purified enzyme in 10 mM Tris, pH 7.0, 2 mM MgCl2, 0.02% NaN3, sitting drop vapour diffusion method, free or in complex with substrate 4-nitrophenyl phosphate, reservoir solution: 12-14% PEG 4000, 20% PEG 3350, 2 mM zinc acetate, 100 mM sodium cacodylate, pH 6.5, the buffer can also be acetate, citrate or imidazole malate, X-ray diffraction structure determination and analysis at 1.9 A resolution
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.1
-
0C, 65% inactivation of the intestinal enzyme, liver enzyme is inactivated at a rate too rapid to be measured, the longer the period in acid the smaller is the percentage of reactivation attainable after returning to pH 7.2
94581
3.5
-
about 50% loss of activity of the liver enzyme after 25 min, about 85% loss of activity of the bone enzyme after 5 min
94582
4.4
-
160 min, about 40% loss of activity of the liver enzyme; about 65% loss of activity of the bone enzyme
94582
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
54
-
20 min, about 45% loss of activity of the liver enzyme, about 60% loss of activity of the bone enzyme
61
-
allozyme D, half-life 10 min, in presence of Zn2+ and Mg2+, pH 9.8
72
-
allozyme S and F, half-life 10 min, in presence of Zn2+ and Mg2+, pH 9.8
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
24 h incubation is necessary for reaching an unfolding equilibrium, dimeric enzyme does not show any sign of dissociation up to a urea concentration of 4 M
-
8 M urea, at 37C, half-life of placental enzyme: 3 h. 3 M urea, half-life of the bone enzyme: 7 min
-
at urea concentrations below a critical value, the enzymes are only slightly and reversibly inhibited, but above this value inactivation is progressive with time and generally irreversible. The critical concentration of urea is considerably greater for intestinal phosphatase than for the kidney and liver enzyme
-
urea, 3 M, 5 h, about 50% loss of activity of the liver enzyme, about 60% loss of activity of the bone enzyme. 5 M, 30 min, about 70% loss of activity of the liver enzyme, about 90% loss of activity of the bone enzyme. 6 M, 10 min, about 70% loss of activity of the liver enzyme, about 90% loss of activity of the bone enzyme
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 18% loss of activity after 2 years
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from placenta
partial purification of isozymes
-
purified from human placental tissue extract. The purity of PLAP after a gel filtration step is above 97%
-
recombinant
-
recombinant enzyme from expression in insect cells
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in COS-1 cell
-
expression in COS-1 cells and CHO K1 Tet-On cells
-
transient expression in COS-1 cell
-
two transgenic rabbit lines are created by pronuclear microinjection with the whey acidic protein promoter-humanTNAP minigene (WAP-hTNAP). lactating females of both lines produce biologically active human TNAP. The recombinant alkaline phosphatase is associated with the membrane of milk fat globules. Use of transgenic rabbits as bioreactors for bulk production of human tissue-nonspecific alkaline phosphatase in milk
allozyme D, ALPP, of placental enzyme form PALP, DNA and amino acid sequence determination of PALP and allozymes, comparison and analysis, substitutions from PALP to ALPP are found at P209R and E429G, subcloning in Escherichia coli XL1-blue, and expression of allozymes D, ALPP, in CHO cells
-
expressed in COS-1 cells
-
expressed in transiently transfected COS-1 cells and in stable CHO-K1 Tet-On cells
-
expression in Pichia pastoris
-
expression in Spodoptera frugiperda Sf9 and Trichoplusia ni BTI-Tn-5B1-4 insect cells via infection with recombinant Autographa california multiple nuclear polyhedrovirus expressing SEAP under control of the polyhedrin promotor, the recombinant enzyme expressed in the 2 different insect cell lines show differing N-glycan structures
-
human 1.6 kb IAP cDNA is subcloned into the pIRES2-EGFP vector, and the construct thus obtained (pIRES2-IAP-EGFP) is used to transfect into IEC-6, T84, and HT-29 cell lines. Stable cell lines are selected by G418 exposure
-
liver and milk isoenzymes have similar or perhaps identical peptide structures and may be coded by the same gene; three gene loci seem to be involved in determining the various forms of alkaline phosphatase, one coding for the placental form, at least one coding for the intestinal forms, and at least one for the liver, bone and kidney forms
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C201Y
-
mutation identified in patient diagnosed with perinatal hypophosphatasia. Mutants exhibit a diminished alkaline phosphatase activity in the cells, where a 66 kDa immature form is predominant with a marginal amount of a 80 kDa mature form. The 66 kDa form exists as a monomer in contrast to a dimer form of wild-type. Only a small fraction of the mutant protein reaches cell surface as the 80 kDa mature form, most of the 66 kDa form is found to be endo-beta-N-acetylglucosaminidase H sensitive and rapidly degraded in proteasome following polyubiquitination
C489S
-
mutation identified in patient diagnosed with perinatal hypophosphatasia. Mutants exhibit a diminished alkaline phosphatase activity in the cells, where a 66 kDa immature form is predominant with a marginal amount of a 80 kDa mature form
G420A
-
naturally occuring mutant, reported in perinatal and childhood hypophosphatasia. Mutant lacks its alkaline phosphatase activity
G420S
-
naturally occuring mutant, reported in perinatal and childhood hypophosphatasia. Mutant lacks its alkaline phosphatase activity, although mutant protein is anchored to the cell surface lipid bilayers by glycosylphosphatidylinositol as an 80 kDa mature form bearing complex-type oligosaccharides similar to wild-type. Mutant G420S largely fails to assemble into the homodimer in contrast to wild-type
N417D
-
mutation abolishes the dimerization without perturbing its cell surface localization
N417E
-
mutation abolishes the dimerization without perturbing its cell surface localization
N417S
-
mutation associated with severe hypophosphatasia, mutants lack enzymic activity. Mutant undergoes N-linked oligosaccharide processing and appears on the cell surface similar to wild-type. Mutant fails to assemble into a dimer structure, which is needed for the catalytic function
A115V
-
activity in U2OS cells after 48h after transfection: 0.1%
A116T
-
mutant responsible for hypophosphatasia shows negligible alkaline phosphatase activity and a weak dominant negative effect when co-expressed with the wild-type enzyme, mutant exists as a monomer and heterogeneously associated aggregates covalently linked via disulfide bonds in contrast to wild-type enzyme which exists as a homodimer
A16V
-
in combination with P275T the mutation causes infantile hypophosphatasia, 7.2% of wild-type activity
M278T
-
lethal mutation, 8.5% of wild-type activity
P275T
-
in combination with A16V mutation causes infantile hypophosphatasia, 4% of wild-type activity
R119H
-
in combination with Y280D the mutation causes childhood hypophosphatasia, 33.4% of wild-type activity
R166A
-
mutation changes the inhibition mechanism of the mutant enzyme to a more complex mixed-type inhibition with decreased affinities for L-Leu and L-Phe
R206W
-
lethal mutation, 2.8% of wild-type activity
R255H
-
mutation (homozygote) causes infantile hypophosphatasia, 6.8% of wild-type activity
R54A
-
activity in U2OS cells after 48h after transfection: 0.1%
T394
-
lethal mutation, 0.3% of wild-type activity
V406A
-
mutant protein shows a markedly reduced alkaline phosphatase activity. Mutant is conveyed to the Golgi apparatus and incorporated into a cold detergent insoluble fraction (raft) at a rate similar to that of the wild-type. Mutant shows increased susceptibility to digestion by trypsin or proteinase K. When purified as a glycosylphosphatidylinositol-anchorless soluble form, mutant protein exhibits a remarkably lower Kcat /Km value compared with that of the wild-type TNSALP
V406F
-
replacement with phenylalanine results in a low enzyme activity, even though TNSALP (V406F) is processed to the 80 kDa mature form similarly to TNSALP (V406A) and appears on the cell surface like the wild-type protein
V406I
-
similar to wild-type, leucine and isoleucine can be successfully substituted for the valine residue
V406L
-
similar to wild-type, leucine and isoleucine can be successfully substituted for the valine residue
Y280D
-
in combination with R119H the mutation causes childhood hypophosphatasia, 1.3% of wild-type activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
medicine
molecular biology