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Literature summary extracted from

  • Truan, D.; Vasil, A.; Stonehouse, M.; Vasil, M.L.; Pohl, E.
    High-level over-expression, purification, and crystallization of a novel phospholipase C/sphingomyelinase from Pseudomonas aeruginosa (2013), Protein Expr. Purif., 90, 40-46.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.4.12 gene plcHR2, expression and secretion of wild-type and L-selenomethionine-labeled enzyme Pseudomonas aeruginosa

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.4.12 purified wild-type enzyme and L-selenomethionine-labeled enzyme, mixing of 0.002 ml of 1.5 mg/ml protein in 50 mM Tris, pH 7.4, 50 mM NaCl, and 0.1 mM TCEP, with 700 nl of reservoir solution containing 10% dioxane, 0.1 M MES, pH 6.75 and 1.5 M ammonium sulfate, and 300 nl 0f 300 mM zwittergent solution, for the labeled enzyme, a protein solution at 9 mg/ml in 50 mM Tris, pH 7.4, 50 mM NaCl, and 0.1 mM TCEP is used, X-ray diffraction structure determination and analysis at 4.0 A and 2.75 A resolution, respectively, molecular replacement Pseudomonas aeruginosa

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.4.12 extracellular in Pseudomonas aeruginosa, the enzyme is co-expressed with the 17 kDa chaperone PlcR2 and is secreted as the fully folded heterodimer PlcHR2, about 95 kDa, by the twin arginine translocase via the cytoplasmic membrane and through the outer membrane, by the Xcp TypeII secretory system Pseudomonas aeruginosa
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Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.4.12 78386
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 2 * 78386, unmodified enzyme, mass spectrometry, 2 * 78500, about, SDS-PAGE, three-dimensional structure analysis of dimeric enzyme Pseudomonas aeruginosa
3.1.4.12 78500
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 2 * 78386, unmodified enzyme, mass spectrometry, 2 * 78500, about, SDS-PAGE, three-dimensional structure analysis of dimeric enzyme Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.4.12 a sphingomyelin + H2O Pseudomonas aeruginosa
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 a ceramide + phosphocholine a ceramide is an N-acylsphingosine ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.4.12 Pseudomonas aeruginosa
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 strain PA01 derivative ADD1976, plcHR2
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Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.4.12 recombinant extracellular wild-type and L-selenomethionine-labeled enzyme from culture supernatant by anion exchange chromatography and gel filtration Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.4.12 a sphingomyelin + H2O
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 Pseudomonas aeruginosa a ceramide + phosphocholine a ceramide is an N-acylsphingosine ?

Subunits

EC Number Subunits Comment Organism
3.1.4.12 dimer 2 * 78386, unmodified enzyme, mass spectrometry, 2 * 78500, about, SDS-PAGE, three-dimensional structure analysis of dimeric enzyme Pseudomonas aeruginosa

Synonyms

EC Number Synonyms Comment Organism
3.1.4.12 phospholipase C/sphingomyelinase
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 Pseudomonas aeruginosa
3.1.4.12 PlcH
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 Pseudomonas aeruginosa

General Information

EC Number General Information Comment Organism
3.1.4.12 physiological function the dimeric enzyme is an important virulence factor in model Pseudomonas aeruginosa infections and is selectively cytotoxic, at picomolar concentrations to mammalian endothelial cells Pseudomonas aeruginosa