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Literature summary for 3.1.3.78 extracted from

  • 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 (2002), EMBO J., 21, 5069-5078.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.3.78

Cloned(Commentary)

Cloned (Comment) 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

Protein Variants

Protein Variants Comment Organism
C438S site-directed mutagenesis Shigella flexneri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphatidylinositol 4,5-bisphosphate + H2O Shigella flexneri
-
 phosphatidylinositol 5-phosphate + phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Shigella flexneri Q07566
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
inositol 1,3,4,5-tetrakisphosphate + H2O
-
 Shigella flexneri inositol 1,3,5-trisphosphate + phosphate
-
 ?
inositol 1,4,5-trisphosphate + H2O
-
 Shigella flexneri inositol 1,5-bisphosphate + phosphate
-
 ?
phosphatidylinositol 3,4,5-trisphosphate + H2O
-
 Shigella flexneri phosphatidylinositol 3,5-bisphosphate + phosphate
-
 ?
phosphatidylinositol 3,4-bisphosphate + H2O
-
 Shigella flexneri phosphatidylinositol 3-phosphate + phosphate
-
 ?
phosphatidylinositol 3-phosphate + H2O
-
 Shigella flexneri phosphatidylinositol + phosphate
-
 ?
phosphatidylinositol 4,5-bisphosphate + H2O
-
 Shigella flexneri phosphatidylinositol 5-phosphate + phosphate
-
 ?
phosphatidylinositol 4-phosphate + H2O
-
 Shigella flexneri phosphatidylinositol + phosphate
-
 ?

Synonyms

Synonyms Comment Organism
IpgD
-
 Shigella flexneri