Literature summary for 3.5.1.23 extracted from

Thayyullathil, F.; Chathoth, S.; Hago, A.; Patel, M.; Szulc, Z.M.; Hannun, Y.; Galadari, S.
Purification and characterization of a second type of neutral ceramidase from rat brain: a second more hydrophobic form of rat brain ceramidase (2011), Biochim. Biophys. Acta, 1811, 242-252.

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Activating Compound

Activating Compound	Comment	Organism	Structure
oxidized glutathione	activity is inhibited almost totally at 20 mM	Rattus norvegicus
reduced glutathione	-	Rattus norvegicus

Inhibitors

Inhibitors	Comment	Organism	Structure
2-mercaptoethanol	more than 50% inhibition at 100 mM	Rattus norvegicus
beta-D-octyl glucoside	almost complete inhibition at 1.5% (v/v)	Rattus norvegicus
Cu2+	about 80% inhibition at 10 mM	Rattus norvegicus
dihydrosphingosine	inhibits enzyme activity by up to 40% at 0.25 mM	Rattus norvegicus
dithiothreitol	80% inhibition at 100 mM	Rattus norvegicus
EDTA	about 60% residual activity at 1 mM	Rattus norvegicus
Fe3+	almost complete inhibition at 10 mM	Rattus norvegicus
Hg2+	complete inhibition at 1 mM	Rattus norvegicus
additional information	GMP, GDT, GTP, TDP, TTP, UDP, and UTP, ADP and AMP have no significant effects on the enzyme activity. S-1-P has no inhibitory effect	Rattus norvegicus
TMP	more than 95% inhibition at 12 mM	Rattus norvegicus
UMP	more than 95% inhibition at 12 mM	Rattus norvegicus
Zn2+	-	Rattus norvegicus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
membrane	-	Rattus norvegicus	16020	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	maximum stimulation at 5 mM Ca2+	Rattus norvegicus
additional information	the enzyme is not affected by Li+, Na+, Mn2+ and Mg2+	Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
95000	-	x * 95000, deglycosylated protein, SDS-PAGE	Rattus norvegicus
110000	-	x * 110000, glycosylated protein, SDS-PAGE	Rattus norvegicus

Organic Solvent Stability

Organic Solvent	Comment	Organism
Triton X-100	the optimum concentration is 0.9-1.2 mM	Rattus norvegicus

Organism

Organism	UniProt	Comment	Textmining
Rattus norvegicus	-	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	-	Rattus norvegicus

Purification (Commentary)

Purification (Comment)	Organism
Q-Sepharose column chromatography, Superdex 200 gel filtration, heparin-Sepharose column chromatography, phenyl-Sepharose column chromatography, and Mono Q column chromatography	Rattus norvegicus

Source Tissue

Source Tissue	Comment	Organism	Textmining
brain	-	Rattus norvegicus	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
1.1	-	non-purified enzyme, at pH 7.5 and 37°C	Rattus norvegicus
16720	-	after 15134fold purification, at pH 7.5 and 37°C	Rattus norvegicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
dihydro-N-myristoyl-D-erythro-sphingosine + H2O	55.9% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	dihydro-D-erythro-sphingosine + myristate	-	?
dihydro-N-palmitoyl-D-erythro-sphingosine + H2O	4.5% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	dihydro-D-erythro-sphingosine + palmitate	-	?
additional information	long chain dihydroceramide dihydro-hexanoyl-D-erythro-sphingosine is resistant to the enzyme	Rattus norvegicus	?	-	?
N-hexanoyl-D-erythro-sphingosine + H2O	100.3% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	sphingosine + hexanoate	-	?
N-lauroyl-D-erythro-sphingosine + H2O	most efficient substrate, 293.2% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	sphingosine + dodecanoate	-	r
N-lignoceroyl-D-erythro-sphingosine + H2O	25.1% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	sphingosine + tetracosanoate	-	?
N-myristoyl-D-erythro-sphingosine + H2O	212.2% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	sphingosine + myristate	-	?
N-palmitoyl-D-erythro-sphingosine + H2O	100% hydrolysis	Rattus norvegicus	sphingosine + palmitate	-	r
N-stearoyl-D-erythro-sphingosine + H2O	98.5% hydrolysis compared to N-palmitoyl-D-erythro-sphingosine	Rattus norvegicus	sphingosine + stearate	-	?

Subunits

Subunits	Comment	Organism
?	x * 110000, glycosylated protein, SDS-PAGE	Rattus norvegicus
?	x * 95000, deglycosylated protein, SDS-PAGE	Rattus norvegicus

Synonyms

Synonyms	Comment	Organism
CDase II	-	Rattus norvegicus
neutral ceramidase	-	Rattus norvegicus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	-	Rattus norvegicus

pI Value

Organism	Comment	pI Value Maximum	pI Value
Rattus norvegicus	isoelectric focusing	-	7.2

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
1	-	in 50 mM Tris (pH 7.5) containing 0.3% (v/v) Triton X-100, at 37°C	Rattus norvegicus	Fe3+
5	-	in 50 mM Tris (pH 7.5) containing 0.3% (v/v) Triton X-100, at 37°C	Rattus norvegicus	Zn2+
20	-	in 50 mM Tris (pH 7.5) containing 0.3% (v/v) Triton X-100, at 37°C	Rattus norvegicus	EDTA

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Release 2023.1 (January 2023)