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Literature summary extracted from

  • Chen, W.; Goldfine, H.; Ananthanarayanan, B.; Cho, W.; Roberts, M.
    Listeria monocytogenes phosphatidylinositol-specific phospholipase C: Kinetic activation and homing in on different interfaces (2009), Biochemistry, 48, 3578-3592.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.4.11 inositol 1,2-(cyclic)-phosphate
-
 Listeria monocytogenes
3.1.4.11 additional information activation by amphiphiles occurs with both micellar (phosphatidylinositol dispersed in detergents) and monomeric [dibutroylphosphatidylinositol (diC4PI)] phosphotransferase substrates and inositol 1,2-(cyclic)-phosphate (cIP), the phosphodiesterase substrate Listeria monocytogenes
4.6.1.13 KCl ionic strength, and not the salt identity, is important for PI-PLC activation towards phosphatidylinositol in micelles. Added salt has a synergistic effect with zwitterionic phospholipids, leading to high specific activities for phosphatidylinositol cleavage with only moderate dilution of the anionic substrate in the interface. This kinetic activation correlates with weakening of strong PI-PLC hydrophobic interactions with the interface. PI-PLC cleavage of phosphatidylinositol presented in small unilamellar vesicles is activated by salt Listeria monocytogenes

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.4.11 transformed into Escherichia coli ER2566 cells for overexpression Listeria monocytogenes
4.6.1.13 inserted into the pTYB11 N-terminal fusion vector between Sap I and EcoR I restriction sites. Recombinant plasmid, IMPACT-Y, transformed into Escherichia coli ER2566 cells for overexpression Listeria monocytogenes

Protein Variants

EC Number Protein Variants Comment Organism
3.1.4.11 F237A site directed mutagenesis Listeria monocytogenes
3.1.4.11 F237W point mutation of surface hydrophobic residues Listeria monocytogenes
3.1.4.11 L151A site directed mutagenesis Listeria monocytogenes
3.1.4.11 L235A point mutation of surface hydrophobic residues Listeria monocytogenes
3.1.4.11 L51A point mutation of surface hydrophobic residues Listeria monocytogenes
3.1.4.11 W49A point mutation of surface hydrophobic residues Listeria monocytogenes
3.1.4.11 W49A/F237A site directed mutagenesis Listeria monocytogenes
4.6.1.13 F237A most approaches wild-type PI-PLC in its dependence on enzyme concentration Listeria monocytogenes
4.6.1.13 F237W even at high concentrations, has high specific activity comparable to dilute unaltered recombinant PI-PLC, does not form the aggregates with anionic lipid-rich vesicles that are disrupted by excess detergent and salt, although it is still activated to about the same extent as wild-type by salt Listeria monocytogenes
4.6.1.13 L151A added KCl (0.15 M) still enhances PI-PLC cleavage of phosphatidylinositol in TX-100 micelles, although KCl effects are much more modest (1.6fold increase) compared to wild-type, F237A or F237W Listeria monocytogenes
4.6.1.13 L235A added KCl (0.15 M) still enhances PI-PLC cleavage of phosphatidylinositol in TX-100 micelles, although KCl effects are much more modest (1.4fold increase) compared to wild-type, F237A or F237W Listeria monocytogenes
4.6.1.13 W49A added KCl (0.15 M) still enhances PI-PLC cleavage of phosphatidylinositol in TX-100 micelles, although KCl effects are much more modest (1.8fold increase) compared to wild-type, F237A or F237W Listeria monocytogenes

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.4.11 12.5
-
 phosphatidylinositol at the lower PI-PLC concentration (3.6 microg/ml) Listeria monocytogenes
3.1.4.11 100
-
 phosphatidylinositol at double the enzyme concentration Listeria monocytogenes

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.4.11 extracellular
-
 Listeria monocytogenes
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.6.1.13 28000
-
 1 * 28000, in solution, laser light scattering Listeria monocytogenes
4.6.1.13 28200
-
 laser light scattering Listeria monocytogenes
4.6.1.13 33000
-
 sequence analysis Listeria monocytogenes

Organism

EC Number Organism UniProt Comment Textmining
3.1.4.11 Listeria monocytogenes A0A0H3GCV8
-
-
4.6.1.13 Listeria monocytogenes
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.4.11 chitin column and SDS-PAGE Listeria monocytogenes
4.6.1.13 by sonication and centrifugation, on chitin column and by gel filtration, more than 85% pure Listeria monocytogenes

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.4.11 additional information
-
 measured using 31P NMR spectroscopy, loss of activity with high enzyme concentrations. Specific activity of the enzyme can be enhanced by dilution of the protein or by addition of both zwitterionic and neutral amphiphiles (e.g., diheptanoylphosphatidylcholine or Triton X-100) or 0.1-0.2 M inorganic salts Listeria monocytogenes
3.1.4.11 0.7
-
 dibutroylphosphatidylinositol as substrate Listeria monocytogenes
3.1.4.11 1.2
-
 dibutroylphosphatidylinositol as substrate, dihexanoylphosphatidylcholine as amphiphile Listeria monocytogenes
3.1.4.11 1.2
-
 dibutroylphosphatidylinositol as substrate, KCl 0.075 M Listeria monocytogenes
3.1.4.11 1.3
-
 phosphatidylinositol as substrate Listeria monocytogenes
3.1.4.11 1.8
-
 dibutroylphosphatidylinositol as substrate, Triton X-100 as amphiphile Listeria monocytogenes
3.1.4.11 9
-
 phosphatidylinositol as substrate, 1-palmitoyl-2-oleoylphosphatidylcholine as amphiphile Listeria monocytogenes
3.1.4.11 10
-
 dibutroylphosphatidylinositol as substrate, dihexanoylphosphatidylcholine as amphiphile, KCl 0.075 M Listeria monocytogenes
3.1.4.11 11.2
-
 dibutroylphosphatidylinositol as substrate, Triton X-100 as amphiphile, KCl 0.075 M Listeria monocytogenes
3.1.4.11 32.7
-
 1.6 microg/ml PI-PLC, citraconic anhydride modification 2 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 54
-
 phosphatidylinositol as substrate, KCl 0.10 M Listeria monocytogenes
3.1.4.11 60.6
-
 1.6 microg/ml PI-PLC, control Listeria monocytogenes
3.1.4.11 82.5
-
 1.6 microg/ml PI-PLC, citraconic anhydride modification 1 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 109
-
 0.2 microg/ml PI-PLC, citraconic anhydride modification 2 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 127
-
 phosphatidylinositol as substrate, 1-palmitoyl-2-oleoylphosphatidylcholine as amphiphile, (NH4)2SO4 0.10 M Listeria monocytogenes
3.1.4.11 165
-
 0.2 microg/ml PI-PLC, citraconic anhydride modification 1 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 274
-
 phosphatidylinositol as substrate, Triton X-100 as amphiphile Listeria monocytogenes
3.1.4.11 303
-
 0.2 microg/ml PI-PLC, control Listeria monocytogenes
3.1.4.11 330
-
 phosphatidylinositol as substrate, KCl 0.20 M Listeria monocytogenes
3.1.4.11 403
-
 0.2 microg/ml PI-PLC, 0.15 M KCl, citraconic anhydride modification 2 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 511
-
 phosphatidylinositol as substrate, KCl 0.30 M Listeria monocytogenes
3.1.4.11 534
-
 0.2 microg/ml PI-PLC, citraconic anhydride modification 2 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 936
-
 phosphatidylinositol as substrate, Triton X-100 as amphiphile, NH4Cl 0.15 M Listeria monocytogenes
3.1.4.11 990
-
 0.2 microg/ml PI-PLC, 0.15 M KCl, citraconic anhydride modification 1 (chemical modification with citraconic anhydride changes the positive charges of lysine residues on the protein surface to negative ones) Listeria monocytogenes
3.1.4.11 1030
-
 0.2 microg/ml PI-PLC, 0.15 M KCl, control Listeria monocytogenes
3.1.4.11 1030
-
 phosphatidylinositol as substrate, Triton X-100 as amphiphile, KCl 0.15 M Listeria monocytogenes
3.1.4.11 1120
-
 phosphatidylinositol as substrate, Triton X-100 as amphiphile, K2SO4 0.10 M Listeria monocytogenes
3.1.4.11 1150
-
 0.2 microg/ml PI-PLC, control Listeria monocytogenes
3.1.4.11 1430
-
 phosphatidylinositol as substrate, Triton X-100 as amphiphile, (NH4)2SO4 0.10 M Listeria monocytogenes

Storage Stability

EC Number Storage Stability Organism
4.6.1.13 4°C, 1 month Listeria monocytogenes

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.4.11 dibutyrylphosphatidylinositol + H2O
-
 Listeria monocytogenes ?
-
 ?
3.1.4.11 additional information water-soluble synthetic short-chain phosphatidylinositol and inositol 1,2-(cyclic)-phosphate along with PI/detergent mixed micelles and PI/PC vesicles are used as substrates Listeria monocytogenes ?
-
 ?
3.1.4.11 phosphatidylinositol + H2O
-
 Listeria monocytogenes diacylglycerol + inositol 1,2-(cyclic)-phosphate
-
 ?
4.6.1.13 dibutyrylphosphatidylinositol is a poor substrate, necessitating long incubation times (2 to 5 h) if the same enzyme concentration is to be used in the absence and presence of salts and amphiphiles Listeria monocytogenes 1D-myo-inositol 1,2-(cyclic)-phosphate + 1,2-dibutyryl-sn-glycerol
-
 ?
4.6.1.13 additional information Phe237 is a key residue in the very tight binding of PI-PLC to membranes containing anionic phospholipids Listeria monocytogenes ?
-
 ?
4.6.1.13 phosphatidylinositol
-
 Listeria monocytogenes diacylglycerol + myo-inositol 1,2-cyclic phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.4.11 monomer in solution in the absence of detergent Listeria monocytogenes
4.6.1.13 monomer 1 * 28000, in solution, laser light scattering Listeria monocytogenes

Synonyms

EC Number Synonyms Comment Organism
3.1.4.11 cyclic phosphodiesterase
-
 Listeria monocytogenes
3.1.4.11 phosphatidylinositol-specific phospholipase
-
 Listeria monocytogenes
3.1.4.11 phosphodiesterase
-
 Listeria monocytogenes
3.1.4.11 PI-PLC
-
 Listeria monocytogenes
3.1.4.11 plcA
-
 Listeria monocytogenes
4.6.1.13 phosphatidylinositol-specific phospholipase C
-
 Listeria monocytogenes
4.6.1.13 PI-PLC
-
 Listeria monocytogenes

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.4.11 7
-
-
 Listeria monocytogenes