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Literature summary for 3.1.4.12 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)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Localization Comment Organism GeneOntology No. Textmining
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
-
-

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa
-
strain PA01 derivative ADD1976, plcHR2
-

Purification (Commentary)

Purification (Comment) Organism
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)

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

Subunits

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

Synonyms

Synonyms Comment Organism
phospholipase C/sphingomyelinase
-
Pseudomonas aeruginosa
PlcH
-
Pseudomonas aeruginosa

General Information

General Information Comment Organism
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