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Information on EC 3.1.3.78 - phosphatidylinositol-4,5-bisphosphate 4-phosphatase
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3.1.3.78 phosphatidylinositol-4,5-bisphosphate 4-phosphataseIUBMB Comments
Two pathways exist in mammalian cells to degrade 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate [PtdIns(4,5)P2] . One is catalysed by this enzyme and the other by EC 3.1.3.36, phosphoinositide 5-phosphatase, where the product is PtdIns4P. The enzyme from human is specific for PtdIns(4,5)P2 as substrate, as it cannot use PtdIns(3,4,5)P3, PtdIns(3,4)P2, PtdIns(3,5)P2, PtdIns5P, PtdIns4P or PtdIns3P . In humans, the enzyme is localized to late endosomal/lysosomal membranes . It can control nuclear levels of PtdIns5P and thereby control p53-dependent apoptosis .
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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
Synonyms
IpgD, phosphatidylinositol-4,5-bisphosphate 4-phosphatase I, phosphatidylinositol-4,5-bisphosphate 4-phosphatase II, phosphoinositide phosphatase, PtdIns(4,5)P2 4-phosphatase, PtdIns-4,5-P2 4-phosphatase type I, PtdIns-4,5-P2 4-phosphatase type II, SigD, type I 4-phosphatase, type I phosphatidylinositol-4,5-bisphosphate 4-phosphatase, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
IpgD
PtdIns-4,5-P2 4-phosphatase type I
-
-
-
-
PtdIns-4,5-P2 4-phosphatase type II
-
-
-
-
SigD
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + H2O = 1-phosphatidyl-1D-myo-inositol 5-phosphate + phosphate
-
-
-
-
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PATHWAY SOURCE
PATHWAYS
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SYSTEMATIC NAME
IUBMB Comments
1-phosphatidyl-1D-myo-inositol-4,5-bisphosphate 4-phosphohydrolase
Two pathways exist in mammalian cells to degrade 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate [PtdIns(4,5)P2] [2]. One is catalysed by this enzyme and the other by EC 3.1.3.36, phosphoinositide 5-phosphatase, where the product is PtdIns4P. The enzyme from human is specific for PtdIns(4,5)P2 as substrate, as it cannot use PtdIns(3,4,5)P3, PtdIns(3,4)P2, PtdIns(3,5)P2, PtdIns5P, PtdIns4P or PtdIns3P [2]. In humans, the enzyme is localized to late endosomal/lysosomal membranes [2]. It can control nuclear levels of PtdIns5P and thereby control p53-dependent apoptosis [3].
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
1D-myo-inositol 1,2,4,5,6-pentakisphosphate + H2O
1D-myo-inositol 1,4,5,6-tetrakisphosphate + phosphate
-
-
-
-
?
D-myo-phosphatidylinositol 4,5-bisphosphate
D-myo-phosphatidylinositol 5-phosphate
-
-
-
-
?
inositol 1,3,4,5-tetrakisphosphate + H2O
inositol 1,3,5-trisphosphate + phosphate
-
-
-
?
inositol 1,4,5-trisphosphate + H2O
inositol 1,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 3,5-bisphosphate + phosphate
phosphatidylinositol 3,4-bisphosphate + H2O
phosphatidylinositol 3-phosphate + phosphate
-
-
-
?
phosphatidylinositol 3-phosphate + H2O
phosphatidylinositol + phosphate
-
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
phosphatidylinositol 5-monophosphate + phosphate
-
-
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
phosphatidylinositol 5-phosphate + phosphate
-
-
-
?
phosphatidylinositol 4-phosphate + H2O
phosphatidylinositol + phosphate
-
-
-
?
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 3,5-bisphosphate + phosphate
-
-
-
-
?
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 3,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 3,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol-4,5-bisphosphate + H2O
phosphatidylinositol-5-phosphate + phosphate
-
-
-
-
?
phosphatidylinositol-4,5-bisphosphate + H2O
phosphatidylinositol-5-phosphate + phosphate
-
-
-
?
additional information
?
-
the enzyme affects epithelial growth factor receptor EGFR degradation upon epithelial growth factor EGF stimulation via phosphatidylinositol-5-phosphate, overview
-
-
?
additional information
?
-
the enzyme affects epithelial growth factor receptor EGFR degradation upon epithelial growth factor EGF stimulation via phosphatidylinositol-5-phosphate, overview
-
-
?
additional information
?
-
-
evidence is provided that SigD functions as a 4â-phosphatase that dephosphorylates D-myo-phosphatidylinositol 4,5-bisphosphate to form D-myo-phosphatidylinositol 5-phosphate
-
-
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 3,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
phosphatidylinositol 5-monophosphate + phosphate
-
-
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
phosphatidylinositol 5-phosphate + phosphate
-
-
-
?
phosphatidylinositol-4,5-bisphosphate + H2O
phosphatidylinositol-5-phosphate + phosphate
-
-
-
?
additional information
?
-
the enzyme affects epithelial growth factor receptor EGFR degradation upon epithelial growth factor EGF stimulation via phosphatidylinositol-5-phosphate, overview
-
-
?
additional information
?
-
the enzyme affects epithelial growth factor receptor EGFR degradation upon epithelial growth factor EGF stimulation via phosphatidylinositol-5-phosphate, overview
-
-
?
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INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
RNAi
-
siRNA of type I 4-phosphatase reduces type I 4-phosphatase levels, which is followed by a dramatic decrease in p53 levels in response to the genotoxic agent etoposide. siRNA of type II 4-phosphatase does not reduce p53 expression
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
etoposide
-
significantly increases type I 4-phosphatase in the nuclear fraction, in both the endogenous and overexpression system
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0000003
-
approx., C462A mutant, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0000005
-
approx., R468A mutant, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0000007
-
approx., GST, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0000011
-
approx., DELTA1-351 mutant, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0000015
-
approx., DELTA352-563 mutant, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0000032
-
approx., DELTA118-142 mutant, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0000063
-
approx., SigD, expressed in Escherichia coli DH5alpha, GST-tagged, malachite-green phosphatase assay (phosphatidylinositol 3,4,5-trisphosphate)
0.0003
10 microM concentration of inositol 1,4,5-trisphosphate and 1500 counts per minute of [3H]inositol 1,4,5-trisphosphate, recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg), reaction 37°C for 10 min in a 50 microl final volume containing 50 mM HEPES pH 7.4, 0.1% bovine serum albumin, 2 mM MgCl2, and 0.033% b-mercaptoethanol, similar results when expressed without IpgE
0.0058
50 microl of recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg) in 50 mM Tris pH 7.5 containing 50 mM glutathione and 100 mM NaCl , 37°C, 15 min, reaction mixture contained 10 microM [32P] phosphatidylinositol 3-phosphate and 20 microM phosphatidylserine, similar results when expressed without IpgE
0.01
50 microl of recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg) in 50 mM Tris pH 7.5 containing 50 mM glutathione and 100 mM NaCl , 37°C, 15 min, reaction mixture contained 10 microM [32P] phosphatidylinositol 4-phosphate and 20 microM phosphatidylserine, similar results when expressed without IpgE
0.0253
50 microl of recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg) in 50 mM Tris pH 7.5 containing 50 mM glutathione and 100 mM NaCl, 37°C, 15 min, reaction mixture contained 10 microM [32P] phosphatidylinositol 3,4-bisphosphate and 20 microM phosphatidylserine, similar results when expressed without IpgE
0.0321
50 microl of recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg) in 50 mM Tris pH 7.5 containing 50 mM glutathione and 100 mM NaCl, 37°C, 15 min, reaction mixture contained 10 microM [32P] phosphatidylinositol 3,4,5-trisphosphate and 20 microM phosphatidylserine, similar results when expressed without IpgE
0.061
50 microl of recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg) in 50 mM Tris pH 7.5 containing 50 mM glutathione and 100 mM NaCl , 37°C, 15 min, reaction mixture contained 10 microM [32P] phosphatidylinositol 4,5-bisphosphate and 20 microM phosphatidylserine, similar results when expressed without IpgE
additional information
not detectable, 1 microM inositol 1,3,4,5-tetrakisphosphate and 1500 counts per minute of [3H]inositol 1,3,4,5-tetrakisphosphate, recombinant GST-IpgD expressed with chaperone IpgE (1-3 microg), reaction 37°C for 10 min in a 50 microl final volume containing 50 mM HEPES pH 7.4, 0.1% bovine serum albumin, 2 mM MgCl2, and 0.033% beta-mercaptoethanol
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
additional information
isozyme II is ubiquitously expressed, comparison of expression patterns of isozymes I and II, overview
brenda
additional information
isozyme II is ubiquitously expressed, comparison of expression patterns of isozymes I and II, overview
brenda
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
when HeLa cells are treated with etoposide or doxorubicin, type I 4-phosphatase translocates to the nucleus and nuclear levels of phosphatidylinositol 5-phosphate increase
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
-
PtdIns(4,5)P2 4-phosphatase expression induces Src kinase and Akt, but not ERK activation and enhances interleukin II promoter activity in T-cells. Expression of a PtdIns5P interacting domain, PH-Dok-5, blocks IpgD-induced T-cell activation and selective signaling molecules downstream of TCR triggering, evaluation, overview
additional information
-
phosphatidylinositol 5-phosphate may play a sensor function in setting the threshold of T-cell activation and contributing to maintain T-cell homeostasis
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C462S
-
no functional or morphological changes in epithelial cells transfected with phosphatase-deficient mutant C462S
C462S
does not restore the ability of Salmonella to eliminate phosphatidylinositol 4,5-bisphosphate in SigD-deficient bacteria
DELTA1-135-DELTA118-142
-
recombinant protein, Saccharomyces cerevisiae: no effect in Saccharomyces cerevisiae, HeLa: no effect (Green Fluorescent Protein linked) in HeLa
DELTA1-165
-
recombinant protein, Saccharomyces cerevisiae: strong impact on actin filaments and budding, HeLa (Green Fluorescent Protein linked): loss of adherence and strong impact on actin filaments
DELTA1-351
-
recombinant protein, Saccharomyces cerevisiae: strong impact on actin filaments and budding, HeLa (Green Fluorescent Protein linked): loss of adherence and strong impact on actin filaments
DELTA118-142
-
recombinant protein, Saccharomyces cerevisiae: strong impact on actin filaments and budding, residues 118â142 crucial for toxicity of SigD in the yeast cells, affecting both phosphatase activity and actin depolarization events, no effect (Green Fluorescent Protein linked) in HeLa
DELTA352-563
-
recombinant protein, no effect in Saccharomyces cerevisiae, no effect in HeLa (Green Fluorescent Protein linked)
K527A
restores the ability of Salmonella to eliminate phosphatidylinositol 4,5-bisphosphate in SigD-deficient bacteria
K530A
restores the ability of Salmonella to eliminate phosphatidylinositol 4,5-bisphosphate in SigD-deficient bacteria
R468A
-
recombinant protein, Saccharomyces cerevisiae: strong impact on actin filaments and budding, HeLa (Green Fluorescent Protein linked): loss of adherence and strong impact on actin filaments
additional information
additional information
expression of isozyme I siRNA in HeLa cells does not affect phosphatidylinositol-4,5-bisphosphate levels or EGFR degradation
additional information
expression of isozyme I siRNA in HeLa cells does not affect phosphatidylinositol-4,5-bisphosphate levels or EGFR degradation
additional information
expression of isozyme II siRNA in HeLa cells does not affect phosphatidylinositol-4,5-bisphosphate levels or EGFR degradation
additional information
expression of isozyme II siRNA in HeLa cells does not affect phosphatidylinositol-4,5-bisphosphate levels or EGFR degradation
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
FLAG-tagged isozyme I from Sf9 insect cells by anti-FLAG immunoaffinity chromatography
FLAG-tagged isozyme II from Sf9 insect cells by anti-FLAG immunoaffinity chromatography
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
(PCR) amplification from the SigD open reading frame from S. enterica ser. typhimurium C53 genomic DNA. 2 types plasmids: first type based on the pEG-KG vector to express GST fusion proteins in yeast, second type based on pGEX-KG for expression of GST-fused proteins in bacteria. transformation of Escherichia coli DH5alpha cells (used for molceularbiological methods and expression). Transformation of Saccharomyces cerevisiae YPH499 (used for SigD functional analyzes as a model organism). transfection of human epithelial cell line HeLa (ATCC CCL2). yeast: endogenous SigD expression results in severe growth inhibition, allele mutant in catalytic site or deletion of whole C-terminal phosphatase domain inhibit growth by loss of actin polarization and precluding the budding process. HeLa: expression of same sigD alleles causes loss of ability of depleting phosphatidylinositol 4,5-bisphosphate from the plasma membrane, actin fibres still disappear and lose their adherence. region of 25 amino acids localized (residues 118â142) essentially required for the effect of SigD on actin in HeLa cells. SigD exerts a toxic effect linked to its N-terminal region and independent of its phosphatase activity
-
by cloning SigD into a mammalian expression vector, it is introduced into intact epithelia cells by microinjection: wild-type SigD induces striking morphological and functional changes that are not mimicked by a phosphatase-deficient SigD mutant C462S
-
infection assay: isotopic equilibrium of Shigella flexneri either wild-type strain M90T, ipgD mutant or the non-invasive strain BS176 (each expressing the AfaE adhesion). Green Fluorescent Protein-tagged IpgD recombinant protein (2 types: wild-type and C438S) cloned in pKN16, transfection of semi-confluent HeLa cells or NIH-3T3 cells. IpgD hydrolyses phosphatidylinositol 4,5-bisphosphate during infection of epithelial cells by Shigella flexneri (analysis of phospholipid content of cells labelled with 32P), phosphatidylinositol 5-phosphate is the product of IpgD-dependent phosphatidylinositol 4,5-bisphosphate degradation in Shigella flexneri infected cells (HPLC, mass assay, immunofluorescence), cellular phosphatidylinositol 4,5-bisphosphate is the only phosphoinositide hydrolysed in vivo, IpgD causes membrane blebbing and cell rounding when expressed in HeLa cells (transiently expressed myc-tagged IpgD in HeLa cells, SDS-PAGE, antibodies against tag or protein), IpgD decreases cytoskeletal-membrane adhesion when expressed in NIH-3T3 cells (optical tweezers)
Salmonella typhimurium SL1344 wild-type and deltaSigD mutant lacking SigD. Transfection of COS-7 and HeLa cells with SigD in mammalian expression vectors. Visualization of phosphatidylinositol 4,5-bisphosphate during Salmonella infection. Fusion protein of PH domain of phospholipase Cdelta and GFP (PLCdelta-PH-GFP) during invasion of HeLA cells by Salmonella (confocal fluorescence microscopy): phosphatidylinositol 4,5-bisphosphate is abundant near the tip but disappears from the basal regions. Role of SigD in phosphatidylinositol 4,5-bisphosphate elimination: focal disappearance of phosphatidylinositol 4,5-bisphosphate observed during invasion (wild-type) is diminished and delayed in the deltaSigD mutant (confocal fluorescence microscopy). SigD alters memebrane elasticity (atomic force microscopy, SigD transfected HeLa cells) and induces vacuole formation. SigD promotes membrane fission during invasion (SigD transfected HeLa cells)
stably transfected HEK-293 TRex cells in a tetracycline-inducible vector expressing type I and II 4-phosphatase
-
type I isozyme, DNA and amino acid sequence determination and analysis, transient transfection of functional enzyme to HeLa and COS-7 cells leading to increased EGFR degradation via increased phosphatidylinositol-5-phosphate content, functional overexpression of FLAG-tagged enzyme in Spodoptera frugiperda Sf9 cells using the baculovirus infection system, stable inducible expression of labeled isozyme I in HEK293 cells
type II isozyme, DNA and amino acid sequence determination and analysis, transient transfection of functional enzyme to HeLa and COS-7 cells leading to increased EGFR degradation via increased phosphatidylinositol-5-phosphate content, functional overexpression of FLAG-tagged enzyme in Spodoptera frugiperda Sf9 cells using the baculovirus infection system
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
medicine
-
involvement of phosphatidylinositol 4,5-bisphosphate phosphatase in Salmonella-induced diarrhea is explored
medicine
-
type I 4-phosphatase regulates nuclear phosphatidylinositol 5-phosphate levels, which in turn mediate p53-dependent apoptosis through interaction with inhibitor of growth protein-2 in response to genotoxic stress
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REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ungewickell, A.; Hugge, C.; Kisseleva, M.; Chang, S.C.; Zou, J.; Feng, Y.; Galyov, E.E.; Wilson, M.; Majerus, P.W.
The identification and characterization of two phosphatidylinositol-4,5-bisphosphate 4-phosphatases
Proc. Natl. Acad. Sci. USA
102
18854-18859
2005
Homo sapiens (Q86T03), Homo sapiens (Q8N4L2)
brenda
Mason, D.; Mallo, G.V.; Terebiznik, M.R.; Payrastre, B.; Finlay, B.B.; Brumell, J.H.; Rameh, L.; Grinstein, S.
Alteration of epithelial structure and function associated with PtdIns(4,5)P2 degradation by a bacterial phosphatase
J. Gen. Physiol.
129
267-283
2007
Salmonella enterica
brenda
Zou, J.; Marjanovic, J.; Kisseleva, M.V.; Wilson, M.; Majerus, P.W.
Type I phosphatidylinositol-4,5-bisphosphate 4-phosphatase regulates stress-induced apoptosis
Proc. Natl. Acad. Sci. USA
104
16834-16839
2007
Homo sapiens
brenda
Aleman, A.; Rodriguez-Escudero, I.; Mallo, G.V.; Cid, V.J.; Molina, M.; Rotger, R.
The amino-terminal non-catalytic region of Salmonella typhimurium SigD affects actin organization in yeast and mammalian cells
Cell. Microbiol.
7
1432-1446
2005
Salmonella enterica subsp. enterica serovar Typhimurium
brenda
Niebuhr, K.; Giuriato, S.; Pedron, T.; Philpott, D.J.; Gaits, F.; Sable, J.; Sheetz, M.P.; Parsot, C.; Sansonetti, P.J.; Payrastre, B.
Conversion of PtdIns(4,5)P(2) into PtdIns(5)P by the S.flexneri effector IpgD reorganizes host cell morphology
EMBO J.
21
5069-5078
2002
Shigella flexneri (Q07566)
brenda
Terebiznik, M.R.; Vieira, O.V.; Marcus, S.L.; Slade, A.; Yip, C.M.; Trimble, W.S.; Meyer, T.; Finlay, B.B.; Grinstein, S.
Elimination of host cell PtdIns(4,5)P(2) by bacterial SigD promotes membrane fission during invasion by Salmonella
Nat. Cell Biol.
4
766-773
2002
Salmonella enterica subsp. enterica serovar Typhimurium (O30916)
brenda
Guittard, G.; Mortier, E.; Tronchere, H.; Firaguay, G.; Gerard, A.; Zimmermann, P.; Payrastre, B.; Nunes, J.A.
Evidence for a positive role of PtdIns5P in T-cell signal transduction pathways
FEBS Lett.
584
2455-2460
2010
Homo sapiens
brenda
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