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Literature summary for 3.5.1.23 extracted from
- Identification of a novel amidase motif in neutral ceramidase (2006), Biochem. J., 393, 687-695.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Saccharomyces cerevisiae enzyme-deficient doubleknockout strain.ypc1.ydc1 microsomes under control of the Gal1 promoter | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C362A | site-directed mutagenesis, inactive enzyme | Homo sapiens |
| D352A | site-directed mutagenesis, inactive enzyme | Homo sapiens |
| S258A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Homo sapiens |
| S354A | site-directed mutagenesis, inactive enzyme | Homo sapiens |
| S374A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
| S396A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Homo sapiens |
| S595A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
| S729A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Homo sapiens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | - |
Homo sapiens |  |
| 2-mercaptoethanol | - |
Rattus norvegicus | |
| Cu2+ | - |
Homo sapiens | |
| Cu2+ | - |
Rattus norvegicus | |
| di-isopropyl fluorophosphate | the substrate protects against inhibition | Homo sapiens | |
| di-isopropyl fluorophosphate | the substrate protects against inhibition | Rattus norvegicus | |
| dithiothreitol | - |
Homo sapiens | |
| dithiothreitol | - |
Rattus norvegicus | |
| Fe2+ | - |
Homo sapiens | |
| Fe2+ | - |
Rattus norvegicus | |
| additional information | no inhibition by protease inhibitors such as aprotinin, PMSF, leupeptin, and pepstatin | Homo sapiens | |
| additional information | no inhibition by protease inhibitors such as aprotinin, PMSF, leupeptin, and pepstatin | Rattus norvegicus | |
| Zn2+ | - |
Homo sapiens | |
| Zn2+ | - |
Rattus norvegicus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | classical MichaelisMenten kinetics | Homo sapiens | |
| 0.06 | - |
D-erythro-C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide | pH 7.5, 37°C, recombinant enzyme | Homo sapiens | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Ca2+ | activates | Rattus norvegicus | |
| Ca2+ | activates | Homo sapiens | |
| Mg2+ | activates slightly | Rattus norvegicus | |
| Mg2+ | activates slightly | Homo sapiens | |
| Mn2+ | activates slightly | Rattus norvegicus | |
| Mn2+ | activates slightly | Homo sapiens | |
| Na+ | activates | Rattus norvegicus | |
| Na+ | activates | Homo sapiens | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | Q9NR71 | - |
- |
| no activity in Saccharomyces cerevisiae | - |
- |
- |
| Rattus norvegicus | Q91XT9 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native enzyme 20000fold from brain by anion exchange chromatography, affinity and hydrophobic interaction chromatography, and cation exchange chromatography, to homogeneity | Rattus norvegicus |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| a ceramide + H2O = a carboxylate + sphingosine | catalytic mechanism | Rattus norvegicus | |
| a ceramide + H2O = a carboxylate + sphingosine | the enzyme has a catalytic residue Ser354 within a very highly conserved hexapeptide sequence GDVSPN in the middle of a larger conserved sequence, namely NXGDVSPNXXGP/XXC, catalytic mechanism | Homo sapiens |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| brain | - |
Rattus norvegicus | - |
| liver | - |
Homo sapiens | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
relative activities of wild-type and mutant enzymes, overview | Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrobenzo-2-oxa-1,3-diazole-C12-ceramide + H2O | - |
Homo sapiens | 4-nitrobenzo-2-oxa-1,3-diazole-dodecanoic acid + ceramide | - |
? | |
| D-erythro-4-nitrobenzo-2-oxa-1,3-diazole-C12-ceramide + H2O | - |
Rattus norvegicus | D-erythro-4-nitrobenzo-2-oxa-1,3-diazole-dodecanoic acid + ceramide | - |
? | |
| D-erythro-4-nitrobenzo-2-oxa-1,3-diazole-C12-ceramide + H2O | - |
Homo sapiens | D-erythro-4-nitrobenzo-2-oxa-1,3-diazole-dodecanoic acid + ceramide | - |
? | |
| D-erythro-C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + H2O | - |
Homo sapiens | D-erythro-sphingosine + ? | - |
? | |
| palmitoyl-sphingosine + H2O | - |
Rattus norvegicus | palmitate + sphingosine | - |
r | |
| palmitoyl-sphingosine + H2O | - |
Homo sapiens | palmitate + sphingosine | - |
r | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CDase | - |
Rattus norvegicus |
| CDase | - |
Homo sapiens |
| neutral ceramidase | - |
Rattus norvegicus |
| neutral ceramidase | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Rattus norvegicus |
| 37 | - |
assay at | Homo sapiens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
- |
Homo sapiens |
| 7.5 | - |
assay at | Rattus norvegicus |
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