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Literature summary extracted from

  • Cheng, J.; Goldstein, R.; Stec, B.; Gershenson, A.; Roberts, M.F.
    Competition between anion binding and dimerization modulates Staphylococcus aureus phosphatidylinositol-specific phospholipase C enzymatic activity (2012), J. Biol. Chem., 287, 40317-40327.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.6.1.13 expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Codonplus (DE3)-RIL Staphylococcus aureus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.6.1.13 purified recombinant wild-type enzyme and mutant Y253S complexed with 1,2-dibutyroyl-sn-glycero-3-phosphocholine, hanging drop vapour diffusion method, mixing of 0.0015 ml of 10 mg/ml protein, 50 mM myo-inositol and 2.5 mM 1,2-dibutyroyl-sn-glycero-3-phosphocholine, with 0.0015 ml of reservoir solution containing 150 mM ammonium acetate, 100 mM sodium acetate, pH 4.6, with 1 mM Mg(NO3)2 (100 mM for the mutant enzymes), and 16-20% PEG 4000, 20°C, 2 weeks, X-ray diffraction structure determination and analysis at 2.16-2.3 A resolution Staphylococcus aureus

Protein Variants

EC Number Protein Variants Comment Organism
4.6.1.13 F249W site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 H86E site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 H86Y site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 V44C site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 V44W site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 Y253K site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 Y253S site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 Y253S/Y255S site-directed mutagenesis, the mutant has the same secondary structure content but a 5°C lower thermal denaturation temperature than the wild-type and an altered enzyme activity Staphylococcus aureus
4.6.1.13 Y253W site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus
4.6.1.13 Y290A site-directed mutagenesis, the mutant shows no significant changes in structure and thermal stability, but altered enzyme activity compared to the wild-type enzyme Staphylococcus aureus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.6.1.13 additional information
-
 additional information phospholipid binding kinetics of the recombinant enzyme, ooverview Staphylococcus aureus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
4.6.1.13 extracellular the enzyme is secreted Staphylococcus aureus
-
-

Organism

EC Number Organism UniProt Comment Textmining
4.6.1.13 Staphylococcus aureus P45723
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.6.1.13 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Codonplus (DE3)-RIL Staphylococcus aureus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.6.1.13 additional information formation of inositol 1,2-(cyclic)phosphate from L-alpha-phoshatidylinositol, NMR spectroscopy analysis Staphylococcus aureus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.6.1.13 dimer the enzyme can form a dimer via helix B, a structural feature present in all secreted, bacterial phosphatidylinositol-specific phospholipases C that is important for membrane binding. The small interface is critical for optimal enzyme activity. The enzyme dimerization is enhanced in membranes containing phosphatidylcholine, fluorescence correlation spectroscopy binding analysis Staphylococcus aureus

Synonyms

EC Number Synonyms Comment Organism
4.6.1.13 More cf. EC 3.1.4.11 Staphylococcus aureus
4.6.1.13 phosphatidylinositol-specific phospholipase C
-
 Staphylococcus aureus
4.6.1.13 PI-PLC
-
 Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.6.1.13 28
-
 assay at Staphylococcus aureus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.6.1.13 6.5
-
-
 Staphylococcus aureus

General Information

EC Number General Information Comment Organism
4.6.1.13 additional information the enzyme can form a dimer via helix B, a structural feature present in all secreted, bacterial phosphatidylinositol-specific phospholipases C that is important for membrane binding. The small interface is critical for optimal enzyme activity. The enzyme dimerization is enhanced in membranes containing phosphatidylcholine, the zwitterionic phospholipid acts not by specific binding to the protein, but rather by reducing anionic lipid interactions with a cationic pocket on the surface of the enzyme that stabilizes monomeric protein. Staphylococcus aureus phosphatidylinositol-specific phospholipase C appears to have a unique mechanism where enzyme activity is modulated by competition between binding of soluble anions or anionic lipids to the cationic sensor and transient dimerization on the membrane Staphylococcus aureus
4.6.1.13 physiological function the enzyme is a virulence factor in the pathogenicity of Staphylococcus aureus Staphylococcus aureus