Cloned (Comment) | Organism |
---|---|
gene plcHR2, expression and secretion of wild-type and L-selenomethionine-labeled enzyme | Pseudomonas aeruginosa |
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 | 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 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas aeruginosa | - |
strain PA01 derivative ADD1976, plcHR2 | - |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a sphingomyelin + H2O | - |
Pseudomonas aeruginosa | a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? |
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 | Comment | Organism |
---|---|---|
phospholipase C/sphingomyelinase | - |
Pseudomonas aeruginosa |
PlcH | - |
Pseudomonas aeruginosa |
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 |