BRENDA - Enzyme Database
show all sequences of 3.1.3.78

Conversion of PtdIns(4,5)P(2) into PtdIns(5)P by the S.flexneri effector IpgD reorganizes host cell morphology

Niebuhr, K.; Giuriato, S.; Pedron, T.; Philpott, D.J.; Gaits, F.; Sable, J.; Sheetz, M.P.; Parsot, C.; Sansonetti, P.J.; Payrastre, B.; EMBO J. 21, 5069-5078 (2002)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
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)
Shigella flexneri
Engineering
Amino acid exchange
Commentary
Organism
C438S
site-directed mutagenesis
Shigella flexneri
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
phosphatidylinositol 4,5-bisphosphate + H2O
Shigella flexneri
-
phosphatidylinositol 5-phosphate + phosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Shigella flexneri
Q07566
-
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
inositol 1,3,4,5-tetrakisphosphate + H2O
-
697606
Shigella flexneri
inositol 1,3,5-trisphosphate + phosphate
-
-
-
?
inositol 1,4,5-trisphosphate + H2O
-
697606
Shigella flexneri
inositol 1,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 3,4,5-trisphosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol 3,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 3,4-bisphosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol 3-phosphate + phosphate
-
-
-
?
phosphatidylinositol 3-phosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol + phosphate
-
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol 5-phosphate + phosphate
-
-
-
?
phosphatidylinositol 4-phosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol + phosphate
-
-
-
?
Cloned(Commentary) (protein specific)
Commentary
Organism
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)
Shigella flexneri
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C438S
site-directed mutagenesis
Shigella flexneri
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
phosphatidylinositol 4,5-bisphosphate + H2O
Shigella flexneri
-
phosphatidylinositol 5-phosphate + phosphate
-
-
?
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
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
Shigella flexneri
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
inositol 1,3,4,5-tetrakisphosphate + H2O
-
697606
Shigella flexneri
inositol 1,3,5-trisphosphate + phosphate
-
-
-
?
inositol 1,4,5-trisphosphate + H2O
-
697606
Shigella flexneri
inositol 1,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 3,4,5-trisphosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol 3,5-bisphosphate + phosphate
-
-
-
?
phosphatidylinositol 3,4-bisphosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol 3-phosphate + phosphate
-
-
-
?
phosphatidylinositol 3-phosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol + phosphate
-
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol 5-phosphate + phosphate
-
-
-
?
phosphatidylinositol 4-phosphate + H2O
-
697606
Shigella flexneri
phosphatidylinositol + phosphate
-
-
-
?
Other publictions for EC 3.1.3.78
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
715020
Guittard
Evidence for a positive role o ...
Homo sapiens
FEBS Lett.
584
2455-2460
2010
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1
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2
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2
2
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681168
Mason
Alteration of epithelial struc ...
Salmonella enterica
J. Gen. Physiol.
129
267-283
2007
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1
1
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1
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1
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3
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1
1
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1
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3
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-
682554
Zou
Type I phosphatidylinositol-4, ...
Homo sapiens
Proc. Natl. Acad. Sci. USA
104
16834-16839
2007
1
1
1
-
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-
2
-
1
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-
2
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1
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1
1
1
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1
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1
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-
666740
Ungewickell
The identification and charact ...
Homo sapiens
Proc. Natl. Acad. Sci. USA
102
18854-18859
2005
-
-
1
-
1
-
-
-
2
-
-
4
-
2
-
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1
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8
-
-
4
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-
2
-
-
2
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-
4
-
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4
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-
2
-
16
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
697201
Aleman
The amino-terminal non-catalyt ...
Salmonella enterica subsp. enterica serovar Typhimurium
Cell. Microbiol.
7
1432-1446
2005
-
-
1
-
7
-
-
-
-
-
-
1
-
1
-
-
-
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7
-
2
-
-
-
-
-
-
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-
1
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7
-
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1
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7
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
697606
Niebuhr
Conversion of PtdIns(4,5)P(2) ...
Shigella flexneri
EMBO J.
21
5069-5078
2002
-
-
1
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
7
-
7
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-
-
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-
-
-
-
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-
-
1
-
-
1
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1
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-
-
-
-
7
-
7
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
700351
Terebiznik
Elimination of host cell PtdIn ...
Salmonella enterica subsp. enterica serovar Typhimurium
Nat. Cell Biol.
4
766-773
2002
-
-
1
-
3
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1
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1
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-
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1
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1
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3
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1
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1
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