BRENDA - Enzyme Database

Purification and characterization of a magnesium-dependent neutral sphingomyelinase from bovine brain

Bernardo, K.; Krut, O.; Wiegmann, K.; Kreder, D.; Micheli, M.; Schafer, R.; Sickman, A.; Schmidt, W.E.; Schroder, J.M.; Meyer, H.E.; Sandhoff, K.; Kronke, M.; J. Biol. Chem. 275, 7641-7647 (2000)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
3.1.4.12
phosphatidylcholine
0.2 mM, 2fold activation
Bos taurus
3.1.4.12
phosphatidylethanolamine
0.2 mM, 2.9fold activation
Bos taurus
3.1.4.12
phosphatidylserine
0.2 mM, 3.8fold activation
Bos taurus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.4.12
Ca2+
0.1 mM, 60% inhibition of Mg2+-activated N-SMase
Bos taurus
3.1.4.12
EDTA
10 mM, 95% inhibition
Bos taurus
3.1.4.12
glutathione
0.2 mM, almost complete inhibition
Bos taurus
3.1.4.12
scyphostatin
0.02 mM, complete inhibition
Bos taurus
3.1.4.12
Zn2+
almost complete inhibition of Mg2+-activated N-SMase
Bos taurus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.4.12
0.04
-
sphingomyelin
37°C, pH 7.4
Bos taurus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.4.12
plasma membrane
associated
Bos taurus
5886
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.1.4.12
Mg2+
required for activity, approx. 3fold activation at 0.1-0.5 mM
Bos taurus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.4.12
97000
-
x * 97000, 2D-electrophoresis
Bos taurus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.1.4.12
sphingomyelin + H2O
Bos taurus
-
N-acylsphingosine + choline phosphate
-
Bos taurus
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.4.12
Bos taurus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.1.4.12
NP-40, CM-Sepharose, concanavalin A-Sepharose, Methyl-macroprep, isoelectric focusing
Bos taurus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.1.4.12
brain
-
Bos taurus
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.1.4.12
0.23
-
-
Bos taurus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.4.12
sphingomyelin + H2O
-
652090
Bos taurus
N-acylsphingosine + choline phosphate
-
652090
Bos taurus
?
Subunits
EC Number
Subunits
Commentary
Organism
3.1.4.12
?
x * 97000, 2D-electrophoresis
Bos taurus
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.1.4.12
56
-
95% loss of activity after 5 min
Bos taurus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.4.12
7
8
-
Bos taurus
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.1.4.12
6
9
approx. 30% of maximal activity at pH 6.5, approx. 70% of maximal activity at pH 9.0
Bos taurus
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
3.1.4.12
Bos taurus
isoelectric focusing
-
5.3
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
3.1.4.12
phosphatidylcholine
0.2 mM, 2fold activation
Bos taurus
3.1.4.12
phosphatidylethanolamine
0.2 mM, 2.9fold activation
Bos taurus
3.1.4.12
phosphatidylserine
0.2 mM, 3.8fold activation
Bos taurus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.4.12
Ca2+
0.1 mM, 60% inhibition of Mg2+-activated N-SMase
Bos taurus
3.1.4.12
EDTA
10 mM, 95% inhibition
Bos taurus
3.1.4.12
glutathione
0.2 mM, almost complete inhibition
Bos taurus
3.1.4.12
scyphostatin
0.02 mM, complete inhibition
Bos taurus
3.1.4.12
Zn2+
almost complete inhibition of Mg2+-activated N-SMase
Bos taurus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.4.12
0.04
-
sphingomyelin
37°C, pH 7.4
Bos taurus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.4.12
plasma membrane
associated
Bos taurus
5886
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.1.4.12
Mg2+
required for activity, approx. 3fold activation at 0.1-0.5 mM
Bos taurus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.4.12
97000
-
x * 97000, 2D-electrophoresis
Bos taurus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.1.4.12
sphingomyelin + H2O
Bos taurus
-
N-acylsphingosine + choline phosphate
-
Bos taurus
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.4.12
NP-40, CM-Sepharose, concanavalin A-Sepharose, Methyl-macroprep, isoelectric focusing
Bos taurus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.1.4.12
brain
-
Bos taurus
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.1.4.12
0.23
-
-
Bos taurus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.4.12
sphingomyelin + H2O
-
652090
Bos taurus
N-acylsphingosine + choline phosphate
-
652090
Bos taurus
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.1.4.12
?
x * 97000, 2D-electrophoresis
Bos taurus
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
3.1.4.12
56
-
95% loss of activity after 5 min
Bos taurus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.4.12
7
8
-
Bos taurus
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
3.1.4.12
6
9
approx. 30% of maximal activity at pH 6.5, approx. 70% of maximal activity at pH 9.0
Bos taurus
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
3.1.4.12
Bos taurus
isoelectric focusing
-
5.3