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Literature summary for 3.5.1.23 extracted from
- Purification, characterization, molecular cloning, and subcellular distribution of neutral ceramidase of rat kidney (2001), J. Biol. Chem., 276, 26249-26259.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| sodium cholate | 112 kDa CDase from kidney, optimum concentration: 0.4-2%, 4-5fold activation | Rattus norvegicus |  |
| sodium taurodeoxycholate | 112 kDa CDase from kidney, optimum concentration: 0.1-0.2%, 4-5fold activation | Rattus norvegicus | |
| Triton X-100 | 2fold activation at 0.1-0.2%, but inhibitory beyond the optimum concentration | Rattus norvegicus | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| 112 kDa-form from kidney, expression in Escherichia coli BL21(DE3), in CHOP cells and overexpression in HEK-239 cells | Rattus norvegicus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Cu2+ | 80% inhibition of 112 kDa CDase from kidney | Rattus norvegicus | |
| Hg2+ | complete inhibition of 112 kDa CDase from kidney | Rattus norvegicus | |
| additional information | EDTA, Mn2+ and Mg2+ have little effect on activity of 112 kDa CDase from kidney | Rattus norvegicus | |
| Triton X-100 | 2fold activation at 0.1-0.2%, but inhibitory beyond the optimum concentration | Rattus norvegicus | |
| Zn2+ | 80% inhibition of 112 kDa CDase from kidney | Rattus norvegicus | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| endoplasmic reticulum | Myc-tagged CDase expressed in HEK-239 cells, a 113 kDa endoplasmic reticulum-form matures into a 133 kDa CDase during processing in the Golgi apparatus | Rattus norvegicus | 5783 | - |
| endosome | CDase is distributed with late endosomes/lysosomes in hepatocytes | Rattus norvegicus | 5768 | - |
| Golgi apparatus | Myc-tagged CDase expressed in HEK-239 cells, a 113 kDa endoplasmic reticulum-form matures into a 133 kDa CDase during processing in the Golgi apparatus | Rattus norvegicus | 5794 | - |
| lysosome | CDase is distributed with late endosomes/lysosomes in hepatocytes | Rattus norvegicus | 5764 | - |
| membrane | membrane-bound, 112 kDa-form, mainly localized at the apical membrane of proximal tubules, distal tubules and collecting ducts of kidney, microsome, sequence contains one possible transmembrane domain | Rattus norvegicus | 16020 | - |
| microsome | kidney, microsomal fraction contains a large amount of membrane-bound 112 kDa neutral CDase | Rattus norvegicus | - |
- |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | 112 kDa CDase from kidney is not activated by Ca2+ | Rattus norvegicus |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 83483 | - |
x * 112000, membrane-bound CDase from kidney, SDS-PAGE, x * 83483, CDase from kidney, sequence calculation, x * 113000, endoplasmic reticulum-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE, x * 133000, Golgi-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE | Rattus norvegicus |
| 112000 | - |
x * 112000, membrane-bound CDase from kidney, SDS-PAGE, x * 83483, CDase from kidney, sequence calculation, x * 113000, endoplasmic reticulum-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE, x * 133000, Golgi-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE | Rattus norvegicus |
| 113000 | - |
x * 112000, membrane-bound CDase from kidney, SDS-PAGE, x * 83483, CDase from kidney, sequence calculation, x * 113000, endoplasmic reticulum-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE, x * 133000, Golgi-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE | Rattus norvegicus |
| 133000 | - |
x * 112000, membrane-bound CDase from kidney, SDS-PAGE, x * 83483, CDase from kidney, sequence calculation, x * 113000, endoplasmic reticulum-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE, x * 133000, Golgi-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE | Rattus norvegicus |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Rattus norvegicus | Q91XT9 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| glycoprotein | 112 kDa CDase from kidney, highly N-glycosylated, sequence contains 9 putative N-glycosylation sites, the N-glycosylation process is indispensable for enzyme activity | Rattus norvegicus |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| 112 kDa-form from kidney: 3604fold, completely insoluble by freeze-thawing in contrast to liver CDase | Rattus norvegicus |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| brain | high CDase expression | Rattus norvegicus | - |
| heart | high CDase expression | Rattus norvegicus | - |
| kidney | high CDase expression, enzyme is mainly localized at the apical membrane of proximal tubules, distal tubules and collecting ducts, enzyme is enriched in the raft microdomains with cholesterol and GM1 ganglioside | Rattus norvegicus | - |
| liver | low CDase expression, enzyme is distributed with late endosomes/lysosomes in hepatocytes | Rattus norvegicus | - |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 1.91 | - |
pH 7.5, 37°C, 112 kDa CDase from kidney, hydrolysis of 4-nitrobenzo-2-oxa-1,3-diazoyl-N-dodecanoylsphingosine | Rattus norvegicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrobenzo-2-oxa-1,3-diazoyl-N-dodecanoylsphingosine + H2O | much faster hydrolysis than of N-lauroylsphingosine | Rattus norvegicus | 4-nitrobenzo-2-oxa-1,3-diazoyl-dodecanoic acid + sphingosine | - |
? | |
| additional information | not: glycosphingolipids, e.g. GalCer, sulfatide, Galbeta(1-3)GalNAcbeta(1-4)(NeuAcalpha(2-3))Galbeta(1-4)Glcbeta(1-1)ceramide, sphingomyelin | Rattus norvegicus | ? | - |
? | |
| N-acylsphingosine + H2O | CDase catalyzes the hydrolysis of the N-acyl linkage of ceramide to produce sphingosine | Rattus norvegicus | a carboxylate + sphingosine | - |
? | |
| N-lauroylsphingosine + H2O | more efficient hydrolysis than of N-palmitoylsphingosine and N-stearoylsphingosine, less efficient hydrolysis than of 4-nitrobenzo-2-oxa-1,3-diazoyl-N-dodecanoylsphingosine | Rattus norvegicus | laurate + sphingosine | - |
? | |
| N-palmitoylsphinganine + H2O | low hydrolysis rate | Rattus norvegicus | palmitate + sphinganine | - |
? | |
| N-palmitoylsphingosine + H2O | less efficient hydrolysis than of N-lauroylsphingosine, more efficient hydrolysis than of N-stearoylsphingosine | Rattus norvegicus | palmitate + sphingosine | - |
? | |
| N-stearoylsphinganine + H2O | low hydrolysis rate | Rattus norvegicus | stearate + sphinganine | - |
? | |
| N-stearoylsphingosine + H2O | less efficient hydrolysis than of N-lauroylsphingosine and N-palmitoylsphingosine | Rattus norvegicus | stearate + sphingosine | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 112000, membrane-bound CDase from kidney, SDS-PAGE, x * 83483, CDase from kidney, sequence calculation, x * 113000, endoplasmic reticulum-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE, x * 133000, Golgi-form of Myc-tagged CDase expressed in HEK-239, SDS-PAGE | Rattus norvegicus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CDase | - |
Rattus norvegicus |
| neutral ceramidase | - |
Rattus norvegicus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Rattus norvegicus |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 7 | 112 kDa CDase from kidney | Rattus norvegicus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 8 | 9 | 112 kDa CDase from kidney, 50% of activity in the range | Rattus norvegicus |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Rattus norvegicus | 112 kDa CDase from kidney, sequence calculation | - |
7.1 |
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