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Literature summary extracted from

  • Galadari, S.; Wu, B.X.; Mao, C.; Roddy, P.; El Bawab, S.; Hannun, Y.A.
    Identification of a novel amidase motif in neutral ceramidase (2006), Biochem. J., 393, 687-695.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.23 expression in Saccharomyces cerevisiae enzyme-deficient doubleknockout strain.ypc1.ydc1 microsomes under control of the Gal1 promoter Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.23 C362A site-directed mutagenesis, inactive enzyme Homo sapiens
3.5.1.23 D352A site-directed mutagenesis, inactive enzyme Homo sapiens
3.5.1.23 S258A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
3.5.1.23 S354A site-directed mutagenesis, inactive enzyme Homo sapiens
3.5.1.23 S374A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
3.5.1.23 S396A site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Homo sapiens
3.5.1.23 S595A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
3.5.1.23 S729A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.23 2-mercaptoethanol
-
 Homo sapiens
3.5.1.23 2-mercaptoethanol
-
 Rattus norvegicus
3.5.1.23 Cu2+
-
 Homo sapiens
3.5.1.23 Cu2+
-
 Rattus norvegicus
3.5.1.23 di-isopropyl fluorophosphate the substrate protects against inhibition Homo sapiens
3.5.1.23 di-isopropyl fluorophosphate the substrate protects against inhibition Rattus norvegicus
3.5.1.23 dithiothreitol
-
 Homo sapiens
3.5.1.23 dithiothreitol
-
 Rattus norvegicus
3.5.1.23 Fe2+
-
 Homo sapiens
3.5.1.23 Fe2+
-
 Rattus norvegicus
3.5.1.23 additional information no inhibition by protease inhibitors such as aprotinin, PMSF, leupeptin, and pepstatin Homo sapiens
3.5.1.23 additional information no inhibition by protease inhibitors such as aprotinin, PMSF, leupeptin, and pepstatin Rattus norvegicus
3.5.1.23 Zn2+
-
 Homo sapiens
3.5.1.23 Zn2+
-
 Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.23 additional information
-
 additional information classical Michaelis–Menten kinetics Homo sapiens
3.5.1.23 0.06
-
 D-erythro-C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide pH 7.5, 37°C, recombinant enzyme Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.23 Ca2+ activates Rattus norvegicus
3.5.1.23 Ca2+ activates Homo sapiens
3.5.1.23 Mg2+ activates slightly Rattus norvegicus
3.5.1.23 Mg2+ activates slightly Homo sapiens
3.5.1.23 Mn2+ activates slightly Rattus norvegicus
3.5.1.23 Mn2+ activates slightly Homo sapiens
3.5.1.23 Na+ activates Rattus norvegicus
3.5.1.23 Na+ activates Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.23 Homo sapiens Q9NR71
-
-
3.5.1.23 no activity in Saccharomyces cerevisiae
-
-
-
3.5.1.23 Rattus norvegicus Q91XT9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.23 native enzyme 20000fold from brain by anion exchange chromatography, affinity and hydrophobic interaction chromatography, and cation exchange chromatography, to homogeneity Rattus norvegicus

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.1.23 a ceramide + H2O = a carboxylate + sphingosine catalytic mechanism Rattus norvegicus
a
3.5.1.23 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
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.5.1.23 brain
-
 Rattus norvegicus
-
3.5.1.23 liver
-
 Homo sapiens
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.1.23 additional information
-
 relative activities of wild-type and mutant enzymes, overview Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.23 4-nitrobenzo-2-oxa-1,3-diazole-C12-ceramide + H2O
-
 Homo sapiens 4-nitrobenzo-2-oxa-1,3-diazole-dodecanoic acid + ceramide
-
 ?
3.5.1.23 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
-
 ?
3.5.1.23 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
-
 ?
3.5.1.23 D-erythro-C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + H2O
-
 Homo sapiens D-erythro-sphingosine + ?
-
 ?
3.5.1.23 palmitoyl-sphingosine + H2O
-
 Rattus norvegicus palmitate + sphingosine
-
 r
3.5.1.23 palmitoyl-sphingosine + H2O
-
 Homo sapiens palmitate + sphingosine
-
 r

Synonyms

EC Number Synonyms Comment Organism
3.5.1.23 CDase
-
 Rattus norvegicus
3.5.1.23 CDase
-
 Homo sapiens
3.5.1.23 neutral ceramidase
-
 Rattus norvegicus
3.5.1.23 neutral ceramidase
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.23 37
-
 assay at Rattus norvegicus
3.5.1.23 37
-
 assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.23 7.5
-
-
 Homo sapiens
3.5.1.23 7.5
-
 assay at Rattus norvegicus