Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | activates in vivo by severalfold | Mus musculus | |
DTT | activates in vivo by severalfold | Mus musculus | |
glutathione | activates in vivo by severalfold | Mus musculus | |
additional information | the enzyme requires a hydrophobic environment for activity | Mus musculus |
Cloned (Comment) | Organism |
---|---|
gene SMPD3, expression of GFP-tagged and of N-terminally RFP-tagged enzyme in HEK-293-IL1R cells, recombinant expression of C-terminally FLAG-tagged enzyme and of enzyme-2-FLAG-6xHis construct in wildtype Escherichia coli strain Rosetta-gami2, and the mutated strain lacking endogenous thioredoxin, results in increased oligomer formation and lower enzyme activity, the phenotype can be rescued by with recombinant human thioredoxin | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
C617S | site-directed mutagenesis, gain-of-function mutation associated with a decreased propensity for oligomerization | Mus musculus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
deoxycholate | inhibits almost 90% of the activity at 3 mM | Mus musculus | |
Diamide | - |
Mus musculus | |
hydrogen peroxide | - |
Mus musculus | |
additional information | oxidizing agents inhibit enzyme activity in a dose-dependent manner that correlates with hyperaggregation of the enzyme protein, which can be restored by immediate addition of DTT | Mus musculus | |
oxidized glutathione | - |
Mus musculus | |
Triton X-100 | inhibits almost 90% of the activity at 0.1% | Mus musculus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | bound | Mus musculus | 16020 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Mus musculus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
74000 | - |
2 * 74000, SDS-PAGE | Mus musculus |
74000 | - |
1 * 74000, SDS-PAGE | Mus musculus |
74000 | - |
3 * 74000, SDS-PAGE | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
a sphingomyelin + H2O | Mus musculus | - |
a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | Q9JJY3 | gene SMPD3 | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme-2-FLAG-6xHis construct from Escherichia coli by affinity chromatography | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a sphingomyelin + H2O | - |
Mus musculus | a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? | |
a sphingomyelin + H2O | 6-N-(7-N´nitro-benz-2-oxa-1,3-diazol-4-yl)aminosphingomyelin is used as substrate | Mus musculus | a ceramide + phosphocholine | a ceramide is an N-acylsphingosine | ? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 74000, SDS-PAGE | Mus musculus |
monomer | 1 * 74000, SDS-PAGE | Mus musculus |
More | the enzyme exists as both monomers and multimers that are associated with high and low enzymatic activity, respectively. Several oxidant-sensitive cysteine residues of the C-terminal catalytic domain are involved in enzyme oligomerization, but cysteines in the non-catalytic region of the enzyme (i.e. amino acids 1 to 337), as well as Cys359, Cys392, Cys420, Cys425, and Cys457 in the beginning of the catalytic domain, do not participate in oligomerization. Thioredoxin-mediated reduction of enzyme multimers to monomers | Mus musculus |
trimer | 3 * 74000, SDS-PAGE | Mus musculus |
Synonyms | Comment | Organism |
---|---|---|
neutral sphingomyelinase-2 | - |
Mus musculus |
nSMase | - |
Mus musculus |
NSMase-2 | - |
Mus musculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Mus musculus |
General Information | Comment | Organism |
---|---|---|
additional information | a redox sensitive enzyme | Mus musculus |
physiological function | neutral sphingomyelinase-2 is the major sphingomyelinase activated in response to proinflammatory cytokines and during oxidative stress. The enzyme may be a novel substrate for thioredoxin | Mus musculus |