Information on EC 3.1.3.67 - phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase

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

EC NUMBER
COMMENTARY hide
3.1.3.67
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RECOMMENDED NAME
GeneOntology No.
phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphate + H2O = 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + phosphate
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
phospho-group transfer
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
3-phosphoinositide degradation
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Inositol phosphate metabolism
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-
SYSTEMATIC NAME
IUBMB Comments
1-phosphatidyl-1D-myo-inositol-3,4,5-trisphosphate 3-phosphohydrolase
Requires Mg2+. Does not dephosphorylate inositol 4,5-bisphosphate. This enzyme still works when the 2,3-bis(acyloxy)propyl group is removed, i.e., it hydrolyses Ins(1,3,4,5)P4 to Ins(1,4,5)P3
CAS REGISTRY NUMBER
COMMENTARY hide
210488-47-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
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phosphatase and tensin homologue is a dual lipidprotein phosphatase that catalyzes the conversion of phosphoinositol 3,4,5-triphosphate to phosphoinositol 4,5-bisphosphate and thereby inhibits PI3K-Akt-dependent cell proliferation, migration, and tumor vascularization. But PTEN is indicated to play a role beyond suppressing PI3K signaling, it also plays a role in regulating Ca2+ entry through transient receptor potential canonical channel 6, TRPC6, that does not require PTEN phosphatase activity, overview. PTEN tail-domain residues 394-403 permit PTEN to associate with TRPC6. The inflammatory mediator thrombin promotes this association. Deletion of PTEN residues 394-403 prevents TRPC6 cell surface expression and Ca2+ entry, regulation, overview
additional information
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loss of PTEN expression in HCT116 and CT26, but not in Caco-2/15, leads to an increase in their metastatic potential following tail-vein injections in mice
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-phosphatidyl-1D-myo-inositol 3,4,5-triphosphate + H2O
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + phosphate
show the reaction diagram
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
show the reaction diagram
-
-
-
?
D-myo-phosphatidylinositol 3,4,5-trisphosphate + H2O
D-myo-phosphatidylinositol 4,5-bisphosphate + phosphate
show the reaction diagram
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-
-
-
?
inositol 1,3,4,5-tetrakisphosphate + H2O
inositol 1,4,5-trisphosphate
show the reaction diagram
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-
-
?
inositol 1,3,4,5-tetrakisphosphate + H2O
inositol 1,4,5-trisphosphate + phosphate
show the reaction diagram
-
-
-
?
phosphatidylinositol 3,4,5-triphosphate + H2O
phosphatidylinositol 4,5-biphosphate + phosphate
show the reaction diagram
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-
-
-
?
phosphatidylinositol 3,4,5-triphosphate + H2O
phosphatidylinositol 4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidyl inositol 4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 4,5-bisphosphate + phosphate
show the reaction diagram
-
-
-
?
phosphatidylinositol 3,4-bisphosphate + H2O
phosphatidylinositol 4-phosphate
show the reaction diagram
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tumor suppressor function. Dephosphorylation of the second-messenger phosphatidylinositol 3,4,5-trisphosphate and phosphatidylinositol 3,4-diphosphate and, by doing so, to antagonize the phosphoinositide 3-kinase pathway
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?
phosphatidylinositol 3,4-bisphosphate + H2O
phosphatidylinositol 4-phosphate + phosphate
show the reaction diagram
phosphatidylinositol 3-phosphate + H2O
phosphatidylinositol + phosphate
show the reaction diagram
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at 20% of the activity with phosphatidylinositol-3,4,5-trisphosphate
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?
Phosphatidylinositol-3,4,5-trisphosphate + H2O
?
show the reaction diagram
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the enzyme may play a critical role in the inositolphospholipid 3-kinase signalling
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-
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Phosphatidylinositol-3,4,5-trisphosphate + H2O
Phosphatidyl inositol-4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol-3,4,5-trisphosphate + H2O
phosphatidylinositol-4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol-3,4-bisphosphate + H2O
phosphatidylinositol 4-phosphate
show the reaction diagram
-
tumor suppressor function. Dephosphorylation of the second-messenger phosphatidylinositol 3,4,5-trisphosphate and phosphatidylinositol 3,4-diphosphate and, by doing so, to antagonize the phosphoinositide 3-kinase pathway
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?
phosphatidylinositol-3,4-bisphosphate + H2O
phosphatidylinositol-4-monophosphate + phosphate
show the reaction diagram
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-
-
?
phosphatidylinostitol 3,4-bisphosphate + H2O
phosphatidylinositol 4-phosphate + phosphate
show the reaction diagram
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at 20% of the activity with phosphatidylinositol-3,4,5-trisphosphate
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?
additional information
?
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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
1-phosphatidyl-1D-myo-inositol 3,4,5-triphosphate + H2O
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + phosphate
show the reaction diagram
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dual-specific phosphatase
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-
?
D-myo-phosphatidylinositol 3,4,5-trisphosphate + H2O
D-myo-phosphatidylinositol 4,5-bisphosphate + phosphate
show the reaction diagram
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-
-
-
?
phosphatidylinositol 3,4,5-triphosphate + H2O
phosphatidylinositol 4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidyl inositol 4,5-bisphosphate + phosphate
show the reaction diagram
Phosphatidylinositol-3,4,5-trisphosphate + H2O
?
show the reaction diagram
-
the enzyme may play a critical role in the inositolphospholipid 3-kinase signalling
-
-
-
Phosphatidylinositol-3,4,5-trisphosphate + H2O
Phosphatidyl inositol-4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol-3,4,5-trisphosphate + H2O
phosphatidylinositol-4,5-bisphosphate + phosphate
show the reaction diagram
phosphatidylinositol-3,4-bisphosphate + H2O
phosphatidylinositol 4-phosphate
show the reaction diagram
-
tumor suppressor function. Dephosphorylation of the second-messenger phosphatidylinositol 3,4,5-trisphosphate and phosphatidylinositol 3,4-diphosphate and, by doing so, to antagonize the phosphoinositide 3-kinase pathway
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?
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
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in presence of 1 mM Mg2+, slight activation up to 0.01 mM, significant inhibition above 0.1 mM
Mg2+
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divalent cation required, maximal activition at 1 mM, inhibition above 1 mM
Mn2+
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divalent cation required, Mn2+ is less effective than Mg2+. 0.5 mM Mn2+ gives 30% of the maximal activity with 1 mM Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-hydroxynonenal
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Ca2+
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in presence of 1 mM Mg2+, slight activation up to 0.01 mM, significant inhibition above 0.1 mM
H2O2
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H2O2 reversibly inhibits PTEN activity by covalently linking Cys-71 to Cys-124, inhibition partially reverses by inclusion of 100 mM dithiothreitol to the buffer
Laminin
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laminin medium induces PTEN upregulation decreases transcription factor Sp1-binding activity
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Mg2+
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divalent cation required, maximal activition at 1 mM, inhibition above 1 mM
phosphatidylserine
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0.2 mM, 75% loss of activity
reactive oxygen species
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oxidation of the active site cysteine by reactive oxygen species inhibits PTEN
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additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acidic lipids
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acidic lipids, and phosphatidylinositol 4,5-bisphosphate in particular, cause a conformational change within the enzyme and allosteric activation
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phosphatidylinositol 4,5-biphosphate
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Enhances PTEN phosphatase activity by inducing conformational change by binding to the PTEN N-terminal domain. Binding constants are in accord with enzyme kinetic measurements showing that phosphatidylinositol 4,5-bisphosphate enhances phosphatidylinositol 3,4,5-triphosphate hydrolysis more effectively than phosphatidylserine, and the binding constant of phosphatidylinositol 4,5-biphosphate of 8.14 microM is in general agreement with the phosphatidylinositol 4,5-bisphosphate concentration requires to activate the PTEN phosphatase. The magnitude of this binding constant indicates that PTEN-phosphatidylinositol 4,5-bisphosphate interactions are physiologically relevant since the effective cellular concentration of phosphatidylinositol 4,5-bisphosphate is thought to be 10 microM, and local phosphatidylinositol 4,5-bisphosphate concentrations are likely to reach even higher levels. Increasing the phosphatidylserine concentration to 25 mol results in stronger binding, but even for these higher surface charge densities, binding constant is still lower than the corresponding values for the phosphatidylcholine/phosphatidylinositol 4,5-bisphosphate vesicle system.
thrombin
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transiently increases PTEN lipid phosphatase activity by about 2.5fold within 1 min, which remains elevated for 5 min
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additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
25.6
4-nitrophenyl phosphate
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0.0989
Inositol 1,3,4,5-tetrakisphosphate
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0.02
phosphatidylinositol 3,4-bisphosphate
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pH not specified in the publication, 23C, Vmax: 0.121 nmol/min/microgram
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12
phosphatidylinositol 3,4-bisphosphate
Ciona intestinalis
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pH not specified in the publication, 23C, Vmax: 0.121 nmol/min/microgram
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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primary astrocyte, expression of wild-type enzyme
Manually annotated by BRENDA team
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cultures are established from wildtype, PTEN heterozygous (+/-) mice and PTEN and Fyn kinase double heterozygous (+/-) mice
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
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decreased enzyme in colorectal cancer
Manually annotated by BRENDA team
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aortic endothelial cells, no change in PTEN protein expression in endothelial cells exposed to up to 24 h of cyclic strain
Manually annotated by BRENDA team
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tissue from patients with sporadic gastric carcinoma
Manually annotated by BRENDA team
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expression of mutant forms of PTEN
Manually annotated by BRENDA team
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human embryonic kidney cell
Manually annotated by BRENDA team
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infiltrative glioma cell line
Manually annotated by BRENDA team
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Madin-darby canine kidney cell
Manually annotated by BRENDA team
amplified by PCR from
Manually annotated by BRENDA team
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the disruption of the PTEN/MMAC1 gene is not a frequent event in neuroblastoma. This disruption may be responsible for malignant progression in only a limited proportion of cases of neuroblastoma
Manually annotated by BRENDA team
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papillomavirus-infected laryngeal papillomas overexpress PTEN/MMAC1
Manually annotated by BRENDA team
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pheochromocytoma cells
Manually annotated by BRENDA team
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derived from testis cDNA
Manually annotated by BRENDA team
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-
Manually annotated by BRENDA team
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gastric cancer cell, PTEN negative and phospho-Akt-positive
Manually annotated by BRENDA team
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glioblastoma-astrocytoma, epithelial-like cell line
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
ectopically expressed GFP-SidF localizes to the ER membrane in mammalian cells. Deletion of the C-terminal portion of SidF including the two transmembrane motifs changes its localization to the cell periphery
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Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513)
Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513)
Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
50-53 kDa, SDS-PAGE
120000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
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1 * 120000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
acetylation
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inhibiton of PTEN
phosphoprotein
ubiquitination
additional information
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carbonylation of PTEN increases significantly in ethanol-fed mice compared to pair-fed control animals, corresponding to decreased PTEN3-phosphatase activity
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures of SidF and its complex with bound short-chain (dibutanoyl) derivative of PI(3,4)P2 is shown. The structures show that the conserved CX5R catalytic motif is located in a large cationic groove
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the 50-amino-acid C-terminal domain, the tail, is necessary for maintaining protein stability
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the PDZ-binding domain binds to several proteins, including MAGI-2 and MAST205, these interactions appear to enhance the stability of PTEN, as interference with either these binding partners or the ability of PTEN to bind them greatly reduces stability. A protein named PICT-1 (protein interacting with the C-tail-1) interacts with the C-terminus of PTEN, promoting both phosphorylation and stability of PTEN
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
fusion-protein is purified by Glutathione-Sepharose-4B affinity chromatography
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Poly-His tag proteins are purified with a HisTrap HP kit using buffers with 10 mM mercaptoethanol, enzymes are further purified with a gel filtration column in 100 mM NaCl, 10 mM Tris, pH 7.4, and 1 mM dithiothreitol. Final purification is done with a anion exchange column in 10 mM Tris, pH 7.4 with a linear gradient from 50-600 mM NaCl.
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recombinant enzyme
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recombinant PTEN is purified to near homogeneity using four sequential column chromatographic steps: a diethylaminoethyl (DEAE) Sepharose anion exchange column, a bio-gel hydroxyapatite HT (HAP) column, a Mono-S cation exchange column, and a Mono-Q anion exchange column; treatment with alkaline phosphatase fully dephosphorylates the phosphorylation sites. Unphosphorylated PTEN and alkaline phosphatase can be separated by ion exchange column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
adenovirus expressing PTEN wild-type, encoding full-length human wild-type PTEN cDNA and AdPTENC/S, encoding a dominant negative human PTEN cDNA mutant (cysteine 124 changed to serine within the catalytic domain) are used for the transduction of cell cultures, overexpression of PTEN and its mutant
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By crossing CD18-Cre recombinase transgenic mice with mice with a conditional point mutation of the pten gene, pten gene is specifically deleted in the B-cell lineage. Mutant animals do not develop B-cell malignancies.; Conditionally deleted pten in oocytes using transgenic mice expressing Cre recombinase under the control of the growth differentiation factor 9 promoter. Pten deficiency in murine oocytes causes the entire oocyte pool to become activated in life.; Creating of a conditional mutant with a combined deletion of Smad4 and Pten, the double mutant shows skin tumor onset; crossing of mice expressing Cre recombinase under the control of the nestin promoter to conditional PTEN mice, mice show a continous increase in brain size throughout embryonal development, individual cells from mutant brain are larger than that of wild type brains; deletion of pten gene in the urothelium results in an increased susceptibility to chemically induced carcinogenesis; female pten-deficient +/- mice develop multifocal endometrial complex atypical hyperplasia between the age of 18 and 39 weeks; Generating of mice with a complete ablation of PTEN is achieved by crossing mice with a conditional point mutation of the pten gene with two transgenic strains in which Cre recmbinase is under the control of the probasin promoter. Mice with complete loss of PTEN in the prostate show 100% penetrance of invasive prostate cancer starting st the age of 6 month; generating of Pten-deficient mice leads to an increasing rate of fatty acid synthesis that is 2.5 times higher than in wild types, furthermore an increase in insulin sensitivity in liver-specifiv Pten-deficient mice is observed, which results in lower fasting plasma glucose levels and reduced serum insulin; Generating of pten +/hyp mice that carry only one hypomorphic pten allele and thus expresses half of the wild type level of one wild type allele. Pten +/hyp mice are crossed with pten +/- mice to generate pten +/+, pten +/-, pten +/hyp and pten hyp/- mice. The pten hyp/- mice are not born at the expected Mendelian ratio, indicating that the hypomorphic pten allele is insufficient to rescue development in all embryos. Surviving male pten hyp/- mice have a much higher incidence of pathological changes in the prostate; Generating of thyroid-specific PTEN-deficient mice using TpoCre transgenic mice,PTEN deletion does not affect normal thyroid development and function, but may contribute to adenoma development; generation of mice with conditional inactivation of PTEN in the mammary epithelium using two different MMTV-Cre recombinase mouse strains results in developmental defects of the mammary gland, mammary ducts in the mutant mice grow much faster than in wild type mice and exhibit excessive side branching and precocious lobuloalveolar budding; homozygous conditional inactivation of PTEN in endothelial and endocardial cells results in embryonic lethality; mice heterozygous for a null mutation of pten gene shows a higher risk for the development of breast and endometrial cancers, by 30-49 weeks of age, 61% female PTEN +/- mice have developed mammary tumors that are mainly adenocarcinomas or small fibroadenomas; mice which deleted pten gene in all T lineage cells show lymphadenopathy, splenomegaly and an enlarged thymus at 6-8 weeks of age, tumor formation is observed from 10 weeks onward and all mice died of malignant T-cell lymphoma by week 17; mice with a conditional null mutation of the pten gene are crossed to transgenic mice in which Cre recombinase expression is driven by the glial fibrillary acidic protein promoter; Mutant mice with brain-specific PTEN deficiency are generated. One transgenic mouse strain expressed Cre recombinase under the control of the engrailed-2 promoter. Crossing of this strain to mice with a conditional null mutation of the pten gene results in the inactivation of PTEN in cells that localize to the dorsal midbrain-hindbrain junction and give rise to cells that populate the vermis of the cerebellum.; Strain of mice with a prostate-specfic deletion of pten is created using Cre recombinase under the control of the prostate-specific antigen promoter, all mutant mice show prostate hyperplasia with focal PIN at the age of 4-5 months. By 7-9 months, PIN is widespread and focal microinvasion is observed. All mutant mice aged 10-14 months show invasive prostate cancer.; the keratin 5 promotor-cren recombinase-driven deletion of pten gene induces hyperplasia of both skin and esophageal squamous epithelium, the esophageal hyperplasia in malnutrition of pups during lactation, 90% of them died within 21 days of birth; To explore PTEN function in the liver, two groups cross PTEN mice containing a conditional point mutation, with AlbCre mice, which express Cre recombinase under the control of albumin promoter. Striking hepatomegaly is observed, which progressed with age. Mutants show accumulation of cytoplasmatic triglycerides that expand over time to severe steatohepatits. In addition, inflammatory cell infiltrates are observed in mutant livers at 24 weeks; transgenic mouse strain expressing Cre recombinase under the control of the L7 promoter, which results in the selective inactivation of PTEN in Purkinje cells
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C-terminal domain of enzyme is expressed in BL21 (DE3) Escherichia coli cells as glutathione S-transferase-protein. Par-3/PDZ3-PTEN peptide single chain fusion protein contains a thrombin cleavage site (LVPRGS) between the C-terminus of PDZ3 domain and the PTEN peptide (DEDQSHQITKV), production of a canine knockdown
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cDNA is subcloned into bicistronic pIRES vector, which also codes for GFP expression and transiently transfected into the following cell lines: HaCaT, MCA3D, NIH 3T3, 3T3 Ki ras and 3T3 v-src
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cytosolic domain (amino acids 215522) is used for analysis
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cytosolic domain (amino acids 248-576) is used for analysis
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expressed in Escherichia coli
expression in Escherichia coli
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GFP-tagged wild-type, C124S and G129E mutant PTEN are constructed
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glioma cell lines U-87MG and U-373MG are stably transfected with wildtype PTEN or catalytically altered mutants of PTEN
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overexpressed in 3T3L1 adipocytes
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overexpressed in MCF-7 breast cancer cell line
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PTEN is ligated into the pBacPAK9 baculovirus transfer vector. Sf9 cells are infected with the recombinant PTEN baculovirus
PTEN liver-specific knock-out mice are generated
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PTEN-null U87 cells are transiently transfected with either wild-type PTEN-green fluorescent protein or PTEN-alanine substituted-green fluorescent protein. U87 cells expressing PH-AKT1-green fluorescent protein are co-transfected with either wildtype PTEN or PTEN-alanine substituted enzyme. Whereas two cells co-expressing wild-type PTEN and PH-AKT1-green fluorescent protein fails to respond to epidermal growth factor stimulation, all PTEN-alanine substituted proteins co-transfected cells fail to respond. HeLa cells are transiently co-transfected with cyan fluorescent protein-FKBP-inositol polyphosphate-5-phosphatase and PTEN-C124S-alanine substituted-yellow fluorescent protein. Expression of PTEN-C124Salanine substituted-yellow fluorescent protein in HeLa cells.
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The cDNA encoding full-length 1-403 PTEN is cloned into the NdeI and XhoI sites of the pET30b vector, thereby introducing a poly-His tag at the C-terminus. Point mutations are introduced with the Quick-Change site-directed mutagenesis kit. Deletion mutant 16-403 is prepared by PCR with the Phusion DNA polymerase. PTEN proteins are expressed in Escherichia coli BL21 (DE3) cells.
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Wild type and mutant cDNA is subcloned in the vector pcDNA3.1 to generate expression vectors, transduction of recombinant PTEN and PTEN mutant into TMK-1 cells. Recombinant PTEN effectively downregulates phospho-Akt levels as well as the LY294002 phosphatidylinositol 3-kinase inhibitor
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
PTEN expression and PTEN phosphorylation are significantly increased in the livers of ethanol-fed mice
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
380-385A
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in endothelial cells transfected with 380-385A (mutation on the PTEN phosphorylation site) phosphatidylinositol 3,4,5-trisphosphate immunofluorescence intensity is than in wildtype or G129R transfected cells. Phosphatidylinositol 3,4,5-trisphosphate intensity in endothelial cells transfected with 380385A is weak but gradually increases to a maximum at 240 min after cyclic strain stimulation by 1.48fold relative to static condition
G129R
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in endothelial cells transfected with G129R, that lacks both protein and lipid phosphatase activities, phosphatidylinositol 3,4,5-trisphosphate immunofluorescence intensity is higher than transfection with wildtype PTEN. phosphatidylinositol 3,4,5-trisphosphate immunofluorescence in endothelial cells transfected with G129R rapidly increases and is maximum at 60 min after cyclic strain stimulation by 2.14fold relative to static condition
DELTA394-403
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removal of the last 10 residues of PTEN leads to the complete diffusion of PTEN in MDCK cells, failing to restore the junctional localization of phosphatidylinositol 4,5-bisphosphate
A121P
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inactive mutant enzyme
C105F
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inactive mutant enzyme
C124R
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inactive mutant enzyme
C136Y
-
inactive mutant enzyme
C71Y
-
inactive mutant enzyme
D107Y
-
inactive mutant enzyme
D331G
-
mutant enzyme with partial activity
D92A
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catalytically inert mutant enzyme, retains partial ability to induce cells to accumulate in G1
DELTA1-15
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phosphatidylinositol 4,5-biphosphate has no effect on binding of PTEN16-403
DELTA352-403
-
truncated enzyme binds strongly to the plasma membrane, an effect that is reversed by co-expression of the remainder of the molecule, PTEN 352-403
F342N
-
mutant enzyme with partial activity
F347L
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mutant enzyme with partial activity
G165R
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inactive mutant enzyme
G20E
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mutant enzyme with partial activity
G251C
-
inactive mutant enzyme
H123Y
-
mutant enzyme lacking phosphatase activity is ineffective in blocking the cell cycle of MCF-7 cells
H129R
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mutant enzyme lacking phosphatase activity is ineffective in blocking the cell cycle of MCF-7 cells
H61R
-
inactive mutant enzyme
H93Y
-
inactive mutant enzyme
K13E
-
the binding of the PTEN mutant K13E, which is a tumor-derived mutation that renders PTEN inactive, is not affected by phosphatidylinositol 4,5-biphosphate
K289E
-
mutant enzyme with partial activity
L112P
-
inactive mutant enzyme
L112R
-
inactive mutant enzyme
L345Q
-
inactive mutant enzyme
L42R
-
activity is comparable with or even higher than that of wild-type enzyme
M134L
-
mutant enzyme with partial activity
PTEN1-274
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when produced as glutathione S-transferase fusion protein, the protein is defective in catalyzing the release of phosphate from either a phosphate-labeled poly(Glu4-Tyr1) substrate or inositol 1,3,4,5-tetrakisphosphate
PTEN1-336
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when produced as glutathione S-transferase fusion protein, the protein is defective in catalyzing the release of phosphate from either a phosphate-labeled poly(Glu4-Tyr1) substrate or inositol 1,3,4,5-tetrakisphosphate
PTENDELTA274-342
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when produced as glutathione S-transferase fusion protein, the protein is defective in catalyzing the release of phosphate from either a phosphate-labeled poly(Glu4-Tyr1) substrate or inositol 1,3,4,5-tetrakisphosphate
R130G
-
inactive mutant enzyme
R130L
-
inactive mutant enzyme
R130Q
-
inactive mutant enzyme
R173C
-
inactive mutant enzyme
R173H
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inactive mutant enzyme
R173P
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inactive mutant enzyme
S10N
-
activity is comparable with or even higher than that of wild-type enzyme
S170N
-
inactive mutant enzyme
S170R
-
inactive mutant enzyme
S227F
-
mutant enzyme with partial activity
S383A
-
PTEN-green fluorescent protein mutant shows significant localization to the plasma membrane, the effect requires no catalytic activity
T383A
-
greater catalytic activity than an unphosphorylated, bacterially expressed wild type enzyme
T385A
-
PTEN-green fluorescent protein mutant shows significant localization to the plasma membrane, the effect requires no catalytic activity
T401I
-
activity is comparable with or even higher than that of wild-type enzyme
V133I
-
inactive mutant enzyme
V343E
-
inactive mutant enzyme
V369G
-
activity is comparable with or even higher than that of wild-type enzyme
Y155C
-
inactive mutant enzyme
Y16C
-
inactive mutant enzyme
Y174N
-
inactive mutant enzyme
Y27S
-
inactive mutant enzyme
Y68H
-
inactive mutant enzyme
C645S
inactive mutant
G129E
-
protein phosphatase-active, lipid phosphatase inactive mutant, expression of G129E mutant of PTEN in U-87MG cells has no effect on the phosphorylation status of protein kinase B, the mutant displays 7% of wildtype lipid phosphatidylinositol 3,4,5-trisphosphate-phosphatase activitiy and 65% wild type protein phosphatase activitiy in glioma cells
R130M
-
kinase dead mutant, expression of R130M mutant of PTEN in U-87MG cells has no effect on the phosphorylation status of protein kinase B, the mutant displays 5% of wildtype lipid phosphatidylinositol 3,4,5-trisphosphate-phosphatase activitiy and 12% of wild-type protein phosphatase activitiy in glioma cells
C124S
-
PTEN mutant is devoid of phosphatase activity and fails to modulate p75NTR expression, indicating that phosphatase activity is required for PTEN regulation of neurotophin receptor p75NTR
G129E
-
PTEN mutant which is lipid phosphatase dead but which retains protein phosphatase activity, significantly reduces the expression of p75NTR, suggesting that it is the protein phosphatase activity of PTEN that is able to regulate neurotophin receptor p75NTR expression
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
availability of the phosphorylated and unphosphorylated forms of recombinant PTEN permits future investigations into the three-dimensional structures of the phosphorylated and unphosphorylated forms of PTEN, and the role of phosphorylation in regulating PTEN activity, phospholipid- and protein-binding affinities
diagnostics
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putative biomarker, nuclear localization of PTEN is important for intestinal differentiation of gastric carcinomas, the loss of cytoplasmatic PTEN expression is correlated with poor gastric carcinoma prognosis
drug development
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This study is a direct evidence for a model in which PTEN switches between open and closed states and phosphorylation favors the closed conformation, thereby regulating localization and function. Small molecules targeting these interactions can potentially serve as therapeutic agents in antagonizing Ras or phosphoinositide 3-kinase-driven tumors.
medicine