Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Bacillus subtilis strain ISW1214 | Bacillus cereus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant N57A enzyme, hanging drop vapor diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH7.0, with 0.002 ml of reservoir solution containing 18% w/v PEG 8000, 0.2 M MgCl2, and 0.1 M sodium cacodylate, pH 6.5, 4°C, X-ray diffraction structure determination and analysis at 2.4 A resolution | Bacillus cereus |
Protein Variants | Comment | Organism |
---|---|---|
D100A | site-directed mutagenesis, mutation in close proximity to Mg2+ at the side-edge, the mutant shows reduced binding to and hydrolysis of sphingomyelin in membranes of sheep erythrocytes or SM-liposomes, similar catalytic activity compared to the wild-type enzyme | Bacillus cereus |
E53A | site-directed mutagenesis, inactive mutant | Bacillus cereus |
E99A | site-directed mutagenesis, mutation in close proximity to Mg2+ at the side-edge, the mutant shows reduced binding to and hydrolysis of sphingomyelin in membranes of sheep erythrocytes or SM-liposomes, similar catalytic activity compared to the wild-type enzyme | Bacillus cereus |
F55A | site-directed mutagenesis, the mutation in close proximity to Mg2+ at the side-edge does not affect the enzyme | Bacillus cereus |
N57A | site-directed mutagenesis, mutation in close proximity to Mg2+ at the side-edge, the mutant shows reduced binding to and hydrolysis of sphingomyelin in membranes of sheep erythrocytes or SM-liposomes, mutant N57A loses the metal ion at the side-edge, similar catalytic activity compared to the wild-type enzyme | Bacillus cereus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | inactivation | Bacillus cereus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | - |
Bacillus cereus | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates | Bacillus cereus | |
Mg2+ | the enzyme is a Mg2+-dependent neutral sphingomyelinase with two metal ion-binding sites in a long horizontal cleft across the molecule, with one Mg2+ in the central region of the cleft and one divalent metal ion at the side-edge of the cleft. The Mg2+ at the side-edge of the enzyme plays an important role in the binding to membranes | Bacillus cereus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
a sphingomyelin + H2O | Bacillus cereus | - |
a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? | |
a sphingomyelin + H2O | Bacillus cereus IAM1029 | - |
a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus cereus | - |
- |
- |
Bacillus cereus IAM1029 | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a sphingomyelin + H2O | - |
Bacillus cereus | a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? | |
a sphingomyelin + H2O | - |
Bacillus cereus IAM1029 | a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? |
Synonyms | Comment | Organism |
---|---|---|
Bc-SMase | - |
Bacillus cereus |
neutral sphingomyelinase | - |
Bacillus cereus |
nSMase | - |
Bacillus cereus |
sphingomyelinase | - |
Bacillus cereus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus cereus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus cereus |