BRENDA - Enzyme Database show
show all sequences of 3.1.3.81

Mutagenesis of the phosphatase sequence motif in diacylglycerol pyrophosphate phosphatase from Saccharomyces cerevisiae

Toke, D.A.; McClintick, M.L.; Carman, G.M.; Biochemistry 38, 14606-14613 (1999)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
gene DPP1, expression of wild-type and phosphatase sequence motif mutant enzymes in an enzyme-deficient mutant yeast strain, subcloning in Escherichia coli strain DH5alpha and XL-1 bLue
Saccharomyces cerevisiae
Engineering
Amino acid exchange
Commentary
Organism
H169A
site-directed mutagenesis of the phosphatase sequence motif residue, the mutant shows 9% of wild-type enzyme DGPP phosphatase activity and no phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
H223A
site-directed mutagenesis of the phosphatase sequence motif residue, the mutant shows 0.03% of wild-type enzyme DGPP phosphatase activity and no phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
R125A
site-directed mutagenesis of the phosphatase sequence motif residue, the mutant shows 0.05% of wild-type enzyme DGPP phosphatase activity and no phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics of recombinant wild-type and mutant enzymes
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
diacylglycerol diphosphate + H2O
Saccharomyces cerevisiae
the substrate and product of the DGPP phosphatase reaction play roles in lipid signaling and in cell metabolism
phosphatidate + phosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Saccharomyces cerevisiae
-
gene DPP1
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0001
-
mutant H223A, DGPP phosphatase activity
Saccharomyces cerevisiae
0.00014
-
mutant R125A, DGPP phosphatase activity
Saccharomyces cerevisiae
0.0269
-
mutant H169A, DGPP phosphatase activity
Saccharomyces cerevisiae
0.0472
-
wild-type enzyme, phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
0.29
-
wild-type enzyme, DGPP phosphatase activity
Saccharomyces cerevisiae
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
diacylglycerol diphosphate + H2O
the substrate and product of the DGPP phosphatase reaction play roles in lipid signaling and in cell metabolism
678120
Saccharomyces cerevisiae
phosphatidate + phosphate
-
-
-
?
diacylglycerol diphosphate + H2O
preferred substrate, the enzyme removes the beta phosphate from DGPP to form phosphatidate
678120
Saccharomyces cerevisiae
phosphatidate + phosphate
-
-
-
?
Subunits
Subunits
Commentary
Organism
More
the primary structure of the DGPP phosphatase protein contains a three-domain phosphatase sequence motif, which is important for catalysis, the residues Arg125, His169, and His223 in domains 1, 2, and 3 are involved
Saccharomyces cerevisiae
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Saccharomyces cerevisiae
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
40
-
10 min, 90% remaing DGPP phosphatase activity of mutant H169A, loss of 20-25% activity of wild-type enzyme and mutants R125A and H223A
Saccharomyces cerevisiae
50
-
10 min, inactivation of mutants R125A and H223A, 80% remaining DGPP phosphatase activity of wild-type enzyme and mutant H169A
Saccharomyces cerevisiae
65
-
10 min, wild-type enzyme and mutant H169A, 75% remaining DGPP phosphatase activity
Saccharomyces cerevisiae
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
assay at
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
Commentary
Organism
gene DPP1, expression of wild-type and phosphatase sequence motif mutant enzymes in an enzyme-deficient mutant yeast strain, subcloning in Escherichia coli strain DH5alpha and XL-1 bLue
Saccharomyces cerevisiae
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
H169A
site-directed mutagenesis of the phosphatase sequence motif residue, the mutant shows 9% of wild-type enzyme DGPP phosphatase activity and no phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
H223A
site-directed mutagenesis of the phosphatase sequence motif residue, the mutant shows 0.03% of wild-type enzyme DGPP phosphatase activity and no phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
R125A
site-directed mutagenesis of the phosphatase sequence motif residue, the mutant shows 0.05% of wild-type enzyme DGPP phosphatase activity and no phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics of recombinant wild-type and mutant enzymes
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
diacylglycerol diphosphate + H2O
Saccharomyces cerevisiae
the substrate and product of the DGPP phosphatase reaction play roles in lipid signaling and in cell metabolism
phosphatidate + phosphate
-
-
?
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.0001
-
mutant H223A, DGPP phosphatase activity
Saccharomyces cerevisiae
0.00014
-
mutant R125A, DGPP phosphatase activity
Saccharomyces cerevisiae
0.0269
-
mutant H169A, DGPP phosphatase activity
Saccharomyces cerevisiae
0.0472
-
wild-type enzyme, phosphatidic acid phosphatase activity
Saccharomyces cerevisiae
0.29
-
wild-type enzyme, DGPP phosphatase activity
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
diacylglycerol diphosphate + H2O
the substrate and product of the DGPP phosphatase reaction play roles in lipid signaling and in cell metabolism
678120
Saccharomyces cerevisiae
phosphatidate + phosphate
-
-
-
?
diacylglycerol diphosphate + H2O
preferred substrate, the enzyme removes the beta phosphate from DGPP to form phosphatidate
678120
Saccharomyces cerevisiae
phosphatidate + phosphate
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
More
the primary structure of the DGPP phosphatase protein contains a three-domain phosphatase sequence motif, which is important for catalysis, the residues Arg125, His169, and His223 in domains 1, 2, and 3 are involved
Saccharomyces cerevisiae
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Saccharomyces cerevisiae
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
40
-
10 min, 90% remaing DGPP phosphatase activity of mutant H169A, loss of 20-25% activity of wild-type enzyme and mutants R125A and H223A
Saccharomyces cerevisiae
50
-
10 min, inactivation of mutants R125A and H223A, 80% remaining DGPP phosphatase activity of wild-type enzyme and mutant H169A
Saccharomyces cerevisiae
65
-
10 min, wild-type enzyme and mutant H169A, 75% remaining DGPP phosphatase activity
Saccharomyces cerevisiae
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6.5
-
assay at
Saccharomyces cerevisiae
Other publictions for EC 3.1.3.81
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
679998
Takeuchi
Cloning and characterization o ...
Homo sapiens
Gene
399
174-180
2007
-
-
1
-
-
-
1
2
-
1
-
-
-
2
-
-
-
-
-
16
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
2
-
2
-
2
-
-
-
-
-
-
-
17
-
-
2
-
2
-
-
-
2
-
-
-
-
-
-
-
-
-
682952
Carman
Roles of phosphatidate phospha ...
Saccharomyces cerevisiae
Trends Biochem. Sci.
31
694-699
2006
-
-
-
-
-
-
3
-
3
1
-
1
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
3
-
-
3
1
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
680628
Han
Vacuole membrane topography of ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae W303-1A
J. Biol. Chem.
279
5338-5345
2004
1
-
1
-
-
-
1
-
3
-
-
2
-
14
-
-
1
1
-
-
2
-
3
1
1
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
1
-
-
3
-
-
2
-
-
-
1
-
-
2
-
3
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
678372
Oshiro
Diacylglycerol pyrophosphate p ...
Saccharomyces cerevisiae
Biochim. Biophys. Acta
1635
1-9
2003
1
-
1
-
4
-
7
-
1
1
1
2
-
1
-
-
1
1
-
-
-
-
3
2
-
-
-
-
1
-
-
-
1
-
-
1
-
1
-
-
4
-
-
7
1
-
1
1
1
2
-
-
-
1
-
-
-
-
3
2
-
-
-
-
1
-
-
-
-
-
-
-
-
-
680624
Oshiro
Regulation of the yeast DPP1-e ...
Saccharomyces cerevisiae
J. Biol. Chem.
278
31495-31503
2003
-
-
1
-
1
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
680613
Han
Regulation of the Saccharomyce ...
Saccharomyces cerevisiae
J. Biol. Chem.
276
10126-10133
2001
-
-
1
-
1
-
1
-
1
-
1
1
-
2
-
-
-
-
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
1
-
-
1
-
-
1
-
1
1
-
-
1
-
1
1
-
-
-
-
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
680612
Oshiro
Regulation of the DPP1-encoded ...
Saccharomyces cerevisiae
J. Biol. Chem.
275
40887-40896
2000
1
-
1
-
-
-
3
-
-
-
1
1
-
1
-
-
-
-
-
-
-
-
2
1
1
-
-
-
1
-
-
-
1
-
1
1
-
1
-
-
-
-
1
3
1
-
-
-
1
1
-
-
-
-
-
-
-
-
2
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
678120
Toke
Mutagenesis of the phosphatase ...
Saccharomyces cerevisiae
Biochemistry
38
14606-14613
1999
-
-
1
-
3
-
-
1
-
-
-
1
-
1
-
-
-
-
-
-
5
-
2
1
1
-
3
-
1
-
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
5
-
2
1
1
-
3
-
1
-
-
-
-
-
-
-
-
-
680606
Toke
Isolation and characterization ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae W303-1A
J. Biol. Chem.
273
3278-3284
1998
-
-
1
-
1
-
-
-
1
1
1
2
-
13
-
-
-
-
-
-
-
-
2
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
1
1
2
-
-
-
-
-
-
-
-
2
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
134878
Carman
Phosphatidate phosphatases and ...
Escherichia coli, Mus musculus, Saccharomyces cerevisiae
Biochim. Biophys. Acta
1348
45-55
1997
-
-
1
-
1
-
5
1
1
4
1
5
-
3
-
-
1
-
-
-
-
-
7
4
-
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
5
-
1
1
4
1
5
-
-
-
1
-
-
-
-
7
4
-
-
-
-
1
-
-
-
-
-
-
-
-
-
680603
Dillon
Mammalian Mg2+-independent pho ...
Rattus norvegicus, Saccharomyces cerevisiae
J. Biol. Chem.
272
10361-10366
1997
1
-
-
-
-
-
6
2
2
2
-
4
-
2
-
-
-
-
-
2
-
-
6
-
2
-
-
-
2
-
-
-
1
-
-
1
-
-
-
-
-
-
-
6
1
2
2
2
-
4
-
-
-
-
-
2
-
-
6
-
2
-
-
-
2
-
-
-
-
-
-
-
-
-
682447
Riedel
-
Metabolism of diacylglycerol p ...
Catharanthus roseus
Plant Sci.
128
1-10
1997
-
-
-
-
-
-
6
-
2
2
-
1
-
1
-
-
-
-
-
4
-
-
3
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
2
2
-
1
-
-
-
-
-
4
-
-
3
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
680601
Wu
Purification and characterizat ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae MATa ade5
J. Biol. Chem.
271
1868-1876
1996
3
-
-
-
-
-
10
1
1
-
1
2
-
2
-
-
1
-
-
-
1
-
4
1
1
-
-
2
1
-
-
-
4
-
-
3
-
-
-
-
-
-
-
10
4
1
1
-
1
2
-
-
-
1
-
-
1
-
4
1
1
-
-
2
1
-
-
-
-
-
-
-
-
-
680602
Dillon
The Escherichia coli pgpB gene ...
Escherichia coli
J. Biol. Chem.
271
30548-30553
1996
1
-
1
-
1
-
2
-
-
1
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
-
1
1
-
1
-
-
1
-
1
2
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-