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Literature summary for 3.1.4.12 extracted from

  • Dotson Ii, P.P.; Karakashian, A.A.; Nikolova-Karakashian, M.N.
    Neutral sphingomyelinase-2 is a redox sensitive enzyme: role of catalytic cysteine residues in regulation of enzymatic activity through changes in oligomeric state (2015), Biochem. J., 465, 371-382.
    View publication on PubMedView publication on EuropePMC

Activating Compound

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(Commentary)

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

Protein Variants Comment Organism
C617S site-directed mutagenesis, gain-of-function mutation associated with a decreased propensity for oligomerization Mus musculus

Inhibitors

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

Localization Comment Organism GeneOntology No. Textmining
membrane bound Mus musculus 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Mus musculus

Molecular Weight [Da]

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Mus musculus Q9JJY3 gene SMPD3
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme-2-FLAG-6xHis construct from Escherichia coli by affinity chromatography Mus musculus

Substrates and Products (Substrate)

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

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

Synonyms Comment Organism
neutral sphingomyelinase-2
-
Mus musculus
nSMase
-
Mus musculus
NSMase-2
-
Mus musculus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Mus musculus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Mus musculus

General Information

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