BRENDA - Enzyme Database

Purification and characterization of diacylglycerol pyrophosphate phosphatase from Saccharomyces cerevisiae

Wu, W.I.; Liu, Y.; Riedel, B.; Wissing, J.B.; Fischl, A.S.; Carman, G.M.; J. Biol. Chem. 271, 1868-1876 (1996)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
3.1.3.81
phosphatidylethanolamine
-
Saccharomyces cerevisiae
3.1.3.81
phosphatidylinositol
-
Saccharomyces cerevisiae
3.1.3.81
Triton X-100
addition of Triton X-100 stimulates DGPP phosphatase activity to a maximum at a concentration of 2 mM
Saccharomyces cerevisiae
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.3.81
4-chloromercuriphenylsulfonic acid
41% inhibition at 1 mM
Saccharomyces cerevisiae
3.1.3.81
ADP
24% inhibition at a 10fold excess of inhibitor to substrate diacylglycerol diphosphate
Saccharomyces cerevisiae
3.1.3.81
Ca2+
-
Saccharomyces cerevisiae
3.1.3.81
diacylglycerol
slight inhibition
Saccharomyces cerevisiae
3.1.3.81
diphosphate
48% inhibition at a 10fold excess of inhibitor to substrate diacylglycerol diphosphate
Saccharomyces cerevisiae
3.1.3.81
Mg2+
-
Saccharomyces cerevisiae
3.1.3.81
Mn2+
-
Saccharomyces cerevisiae
3.1.3.81
additional information
poor inhibition by ATP and AMP, the enzyme is inhibited by divalent cations, but is relatively insensitive to thioreactive agents, no inhibition by phospholipids or phosphatidic acid versus diacylglycerol diphosphate as substrate, no inhibition by EDTA, glycerol 3-phosphate, and inositol 1-phosphate
Saccharomyces cerevisiae
3.1.3.81
NaF
94% inhibition at 10 mM
Saccharomyces cerevisiae
3.1.3.81
NEM
10% inhibition at 5 mM
Saccharomyces cerevisiae
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.3.81
additional information
-
additional information
kinetics, the DGPP phosphatase exhibits typical saturation kinetics with respect to DGPP, the Km value for diacylglycerol diphosphate is 3fold greater than its cellular concentration of 0.18 mol%
Saccharomyces cerevisiae
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.3.81
membrane
associated
Saccharomyces cerevisiae
16020
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.3.81
34000
-
x * 34000, SDS-PAGE
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.1.3.81
diacylglycerol diphosphate + H2O
Saccharomyces cerevisiae
preferred substrate
phosphatidate + phosphate
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
Saccharomyces cerevisiae MATa ade5
preferred substrate
phosphatidate + phosphate
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.1.3.81
Saccharomyces cerevisiae
-
-
-
3.1.3.81
Saccharomyces cerevisiae MATa ade5
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.1.3.81
33333fold to homogeneity by solubilization from membranes with Triton X-100, followed by anion exchange chromatography, affinity and hydroxylapatite chromatography, and again anion exchange chromatography
Saccharomyces cerevisiae
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.1.3.81
150
-
purified enzyme
Saccharomyces cerevisiae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate
680601
Saccharomyces cerevisiae
phosphatidate + phosphate
-
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate, the bifunctional DPP1 catalyzes the removal of the beta-phosphate from diacylglycerol diphosphate to form phosphatidate, and it also removes the phosphate from phosphatidate to form diacylglycerol, reaction of EC 3.1.3.4
680601
Saccharomyces cerevisiae
phosphatidate + phosphate
product identification by TLC
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate
680601
Saccharomyces cerevisiae MATa ade5
phosphatidate + phosphate
-
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate, the bifunctional DPP1 catalyzes the removal of the beta-phosphate from diacylglycerol diphosphate to form phosphatidate, and it also removes the phosphate from phosphatidate to form diacylglycerol, reaction of EC 3.1.3.4
680601
Saccharomyces cerevisiae MATa ade5
phosphatidate + phosphate
product identification by TLC
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.1.3.81
?
x * 34000, SDS-PAGE
Saccharomyces cerevisiae
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.1.3.81
30
-
assay at
Saccharomyces cerevisiae
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.3.81
40.9
-
diacylglycerol diphosphate
pH 6.5, 30°C
Saccharomyces cerevisiae
3.1.3.81
96.67
-
diacylglycerol diphosphate
pH 6.5, 30°C
Saccharomyces cerevisiae
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.3.81
6
8.5
-
Saccharomyces cerevisiae
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.1.3.81
additional information
-
additional information
-
Saccharomyces cerevisiae
3.1.3.81
0.058
-
Mn2+
pH 6.5, 30°C
Saccharomyces cerevisiae
3.1.3.81
0.56
-
Ca2+
pH 6.5, 30°C
Saccharomyces cerevisiae
3.1.3.81
17
-
Mg2+
pH 6.5, 30°C
Saccharomyces cerevisiae
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
3.1.3.81
phosphatidylethanolamine
-
Saccharomyces cerevisiae
3.1.3.81
phosphatidylinositol
-
Saccharomyces cerevisiae
3.1.3.81
Triton X-100
addition of Triton X-100 stimulates DGPP phosphatase activity to a maximum at a concentration of 2 mM
Saccharomyces cerevisiae
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.1.3.81
4-chloromercuriphenylsulfonic acid
41% inhibition at 1 mM
Saccharomyces cerevisiae
3.1.3.81
ADP
24% inhibition at a 10fold excess of inhibitor to substrate diacylglycerol diphosphate
Saccharomyces cerevisiae
3.1.3.81
Ca2+
-
Saccharomyces cerevisiae
3.1.3.81
diacylglycerol
slight inhibition
Saccharomyces cerevisiae
3.1.3.81
diphosphate
48% inhibition at a 10fold excess of inhibitor to substrate diacylglycerol diphosphate
Saccharomyces cerevisiae
3.1.3.81
Mg2+
-
Saccharomyces cerevisiae
3.1.3.81
Mn2+
-
Saccharomyces cerevisiae
3.1.3.81
additional information
poor inhibition by ATP and AMP, the enzyme is inhibited by divalent cations, but is relatively insensitive to thioreactive agents, no inhibition by phospholipids or phosphatidic acid versus diacylglycerol diphosphate as substrate, no inhibition by EDTA, glycerol 3-phosphate, and inositol 1-phosphate
Saccharomyces cerevisiae
3.1.3.81
NaF
94% inhibition at 10 mM
Saccharomyces cerevisiae
3.1.3.81
NEM
10% inhibition at 5 mM
Saccharomyces cerevisiae
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
3.1.3.81
additional information
-
additional information
-
Saccharomyces cerevisiae
3.1.3.81
0.058
-
Mn2+
pH 6.5, 30°C
Saccharomyces cerevisiae
3.1.3.81
0.56
-
Ca2+
pH 6.5, 30°C
Saccharomyces cerevisiae
3.1.3.81
17
-
Mg2+
pH 6.5, 30°C
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.1.3.81
additional information
-
additional information
kinetics, the DGPP phosphatase exhibits typical saturation kinetics with respect to DGPP, the Km value for diacylglycerol diphosphate is 3fold greater than its cellular concentration of 0.18 mol%
Saccharomyces cerevisiae
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.1.3.81
membrane
associated
Saccharomyces cerevisiae
16020
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.3.81
34000
-
x * 34000, SDS-PAGE
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3.1.3.81
diacylglycerol diphosphate + H2O
Saccharomyces cerevisiae
preferred substrate
phosphatidate + phosphate
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
Saccharomyces cerevisiae MATa ade5
preferred substrate
phosphatidate + phosphate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.3.81
33333fold to homogeneity by solubilization from membranes with Triton X-100, followed by anion exchange chromatography, affinity and hydroxylapatite chromatography, and again anion exchange chromatography
Saccharomyces cerevisiae
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.1.3.81
150
-
purified enzyme
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate
680601
Saccharomyces cerevisiae
phosphatidate + phosphate
-
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate, the bifunctional DPP1 catalyzes the removal of the beta-phosphate from diacylglycerol diphosphate to form phosphatidate, and it also removes the phosphate from phosphatidate to form diacylglycerol, reaction of EC 3.1.3.4
680601
Saccharomyces cerevisiae
phosphatidate + phosphate
product identification by TLC
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate
680601
Saccharomyces cerevisiae MATa ade5
phosphatidate + phosphate
-
-
-
?
3.1.3.81
diacylglycerol diphosphate + H2O
preferred substrate, the bifunctional DPP1 catalyzes the removal of the beta-phosphate from diacylglycerol diphosphate to form phosphatidate, and it also removes the phosphate from phosphatidate to form diacylglycerol, reaction of EC 3.1.3.4
680601
Saccharomyces cerevisiae MATa ade5
phosphatidate + phosphate
product identification by TLC
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.1.3.81
?
x * 34000, SDS-PAGE
Saccharomyces cerevisiae
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.1.3.81
30
-
assay at
Saccharomyces cerevisiae
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.1.3.81
40.9
-
diacylglycerol diphosphate
pH 6.5, 30°C
Saccharomyces cerevisiae
3.1.3.81
96.67
-
diacylglycerol diphosphate
pH 6.5, 30°C
Saccharomyces cerevisiae
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.3.81
6
8.5
-
Saccharomyces cerevisiae