BRENDA - Enzyme Database
show all sequences of 2.7.1.174

Characterization of the yeast DGK1-encoded CTP-dependent diacylglycerol kinase

Han, G.S.; OHara, L.; Siniossoglou, S.; Carman, G.M.; J. Biol. Chem. 283, 20443-20453 (2008)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
dolichol
-
Saccharomyces cerevisiae
additional information
diacylglycerol kinase activity is stimulated by major membrane phospholipids, overview
Saccharomyces cerevisiae
phosphatidate
-
Saccharomyces cerevisiae
phosphatidylcholine
-
Saccharomyces cerevisiae
phosphatidylethanolamine
-
Saccharomyces cerevisiae
phosphatidylglycerol
-
Saccharomyces cerevisiae
phosphatidylinositol
-
Saccharomyces cerevisiae
phosphatidylserine
-
Saccharomyces cerevisiae
Cloned(Commentary)
Commentary
Organism
gene DGK1, the DGK1 promoter is substituted with the inducible GAL1/10 promoter in the low copy YCplac111 and high copy YEplac181 vectors, expression of wild-type and mutant enzymes, induction of wild-type DGK1 gene expression from high copy number plasmid YEplac181-GAL1/10-DGK1 results in a massive increase in DAG kinase activity. Temperature sensitivity of dgk1DELTA cells overexpressing DGK1 and its mutant alleles
Saccharomyces cerevisiae
Engineering
Amino acid exchange
Commentary
Organism
D177A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
G184A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
K77A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing an almost complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
additional information
construction of DGK1 truncation mutants. The DELTA66 and DELTA70 truncations remove most of the N-terminal hydrophilic region of Dgk1p, whereas the DELTA77 truncation removes the entire N-terminal hydrophilic region plus the first two residues contained within the CTP transferase domain, the later mutant is inactive, the other two show reduced activity compared to the wild-type enzyme
Saccharomyces cerevisiae
R76A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
Inhibitors
Inhibitors
Commentary
Organism
Structure
cardiolipin
-
Saccharomyces cerevisiae
CDP-diacylglycerol
-
Saccharomyces cerevisiae
ceramide
-
Saccharomyces cerevisiae
dCTP
both a substrate and competitive inhibitor
Saccharomyces cerevisiae
diacylglycerol diphosphate
-
Saccharomyces cerevisiae
lyso-phosphatidate
-
Saccharomyces cerevisiae
Mn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
additional information
the enzyme is inhibited by sphingoid bases
Saccharomyces cerevisiae
N-ethylmaleimide
-
Saccharomyces cerevisiae
phytosphingosine
-
Saccharomyces cerevisiae
R59022
25% inhibition at 0.05 M
Saccharomyces cerevisiae
R59949
15% inhibition at 0.05 M
Saccharomyces cerevisiae
sphinganine
-
Saccharomyces cerevisiae
sphingosine
-
Saccharomyces cerevisiae
triacylglycerol
-
Saccharomyces cerevisiae
Triton X-100
inhibition kinetics, overview. No inhibition by monoacylglycerol
Saccharomyces cerevisiae
Zn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
the enzyme exhibits positive cooperative kinetics with respect to diacylglycerol and saturation kinetics with respect to CTP, kinetic analysis and modeling, overview
Saccharomyces cerevisiae
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
-
Saccharomyces cerevisiae
16020
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Ca2+
the enzyme requires Ca2+ or Mg2+ ions for activity. Maximum DAG kinase activity obtained with 1 mM Ca2+ ions is slightly greater than the maximum activity obtained with 10 mM Mg2+ ions
Saccharomyces cerevisiae
Mg2+
the enzyme requires Ca2+ or Mg2+ ions for activity. Maximum DAG kinase activity obtained with 1 mM Ca2+ ions is slightly greater than the maximum activity obtained with 10 mM Mg2+ ions
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
CTP + 1,2-diacyl-sn-glycerol
Saccharomyces cerevisiae
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
?
CTP + 1,2-diacyl-sn-glycerol
Saccharomyces cerevisiae BY4742
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Saccharomyces cerevisiae
Q12382
gene DGK1 or YOR311C
-
Saccharomyces cerevisiae BY4742
Q12382
gene DGK1 or YOR311C
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.018
-
wild-type cells, pH 7.5, 30°C
Saccharomyces cerevisiae
130
-
DAG kinase in the membrane fraction of galactose-grown cells, pH 7.5, 30°C
Saccharomyces cerevisiae
Storage Stability
Storage Stability
Organism
-80°C, the enzyme preparation is completely stable for at least 3 months of storage at, and is stable to several cycles of freezing and thawing
Saccharomyces cerevisiae
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
CTP + 1,2-diacyl-sn-glycerol
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
687738
Saccharomyces cerevisiae
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
CTP + 1,2-diacyl-sn-glycerol
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
687738
Saccharomyces cerevisiae BY4742
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
CTP + 1,2-dioleoyl-sn-glycerol
the CTP transferase domain is sufficient for diacylglycerol kinase activity
687738
Saccharomyces cerevisiae
CDP + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
-
-
?
CTP + 1,2-dioleoyl-sn-glycerol
the CTP transferase domain is sufficient for diacylglycerol kinase activity
687738
Saccharomyces cerevisiae BY4742
CDP + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
-
-
?
dCTP + 1,2-diacyl-sn-glycerol
both a substrate and competitive inhibitor
687738
Saccharomyces cerevisiae
dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
dCTP + 1,2-diacyl-sn-glycerol
both a substrate and competitive inhibitor
687738
Saccharomyces cerevisiae BY4742
dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
Subunits
Subunits
Commentary
Organism
More
Dgk1p contains a CTP transferase domain, the CTP transferase domain is sufficient for diacylglycerol kinase activity
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
-
stable up to, labile above
Saccharomyces cerevisiae
70
-
loss of about 70% of the activity
Saccharomyces cerevisiae
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
7.5
-
Saccharomyces cerevisiae
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6.5
9
activity range, profile overview
Saccharomyces cerevisiae
Cofactor
Cofactor
Commentary
Organism
Structure
CTP
dependent on
Saccharomyces cerevisiae
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
additional information
-
additional information
surface dilution kinetic model, overview, positive cooperative kinetics with respect to the surface concentration of 1,2-dioleoyl-sn-glycerol
Saccharomyces cerevisiae
IC50 Value
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.1
-
pH 7.5, 30°C
Saccharomyces cerevisiae
N-ethylmaleimide
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
dolichol
-
Saccharomyces cerevisiae
additional information
diacylglycerol kinase activity is stimulated by major membrane phospholipids, overview
Saccharomyces cerevisiae
phosphatidate
-
Saccharomyces cerevisiae
phosphatidylcholine
-
Saccharomyces cerevisiae
phosphatidylethanolamine
-
Saccharomyces cerevisiae
phosphatidylglycerol
-
Saccharomyces cerevisiae
phosphatidylinositol
-
Saccharomyces cerevisiae
phosphatidylserine
-
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
Commentary
Organism
gene DGK1, the DGK1 promoter is substituted with the inducible GAL1/10 promoter in the low copy YCplac111 and high copy YEplac181 vectors, expression of wild-type and mutant enzymes, induction of wild-type DGK1 gene expression from high copy number plasmid YEplac181-GAL1/10-DGK1 results in a massive increase in DAG kinase activity. Temperature sensitivity of dgk1DELTA cells overexpressing DGK1 and its mutant alleles
Saccharomyces cerevisiae
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
CTP
dependent on
Saccharomyces cerevisiae
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D177A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
G184A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
K77A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing an almost complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
additional information
construction of DGK1 truncation mutants. The DELTA66 and DELTA70 truncations remove most of the N-terminal hydrophilic region of Dgk1p, whereas the DELTA77 truncation removes the entire N-terminal hydrophilic region plus the first two residues contained within the CTP transferase domain, the later mutant is inactive, the other two show reduced activity compared to the wild-type enzyme
Saccharomyces cerevisiae
R76A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
IC50 Value (protein specific)
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
0.1
-
pH 7.5, 30°C
Saccharomyces cerevisiae
N-ethylmaleimide
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
cardiolipin
-
Saccharomyces cerevisiae
CDP-diacylglycerol
-
Saccharomyces cerevisiae
ceramide
-
Saccharomyces cerevisiae
dCTP
both a substrate and competitive inhibitor
Saccharomyces cerevisiae
diacylglycerol diphosphate
-
Saccharomyces cerevisiae
lyso-phosphatidate
-
Saccharomyces cerevisiae
Mn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
additional information
the enzyme is inhibited by sphingoid bases
Saccharomyces cerevisiae
N-ethylmaleimide
-
Saccharomyces cerevisiae
phytosphingosine
-
Saccharomyces cerevisiae
R59022
25% inhibition at 0.05 M
Saccharomyces cerevisiae
R59949
15% inhibition at 0.05 M
Saccharomyces cerevisiae
sphinganine
-
Saccharomyces cerevisiae
sphingosine
-
Saccharomyces cerevisiae
triacylglycerol
-
Saccharomyces cerevisiae
Triton X-100
inhibition kinetics, overview. No inhibition by monoacylglycerol
Saccharomyces cerevisiae
Zn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
additional information
-
additional information
surface dilution kinetic model, overview, positive cooperative kinetics with respect to the surface concentration of 1,2-dioleoyl-sn-glycerol
Saccharomyces cerevisiae
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
the enzyme exhibits positive cooperative kinetics with respect to diacylglycerol and saturation kinetics with respect to CTP, kinetic analysis and modeling, overview
Saccharomyces cerevisiae
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
-
Saccharomyces cerevisiae
16020
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Ca2+
the enzyme requires Ca2+ or Mg2+ ions for activity. Maximum DAG kinase activity obtained with 1 mM Ca2+ ions is slightly greater than the maximum activity obtained with 10 mM Mg2+ ions
Saccharomyces cerevisiae
Mg2+
the enzyme requires Ca2+ or Mg2+ ions for activity. Maximum DAG kinase activity obtained with 1 mM Ca2+ ions is slightly greater than the maximum activity obtained with 10 mM Mg2+ ions
Saccharomyces cerevisiae
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
CTP + 1,2-diacyl-sn-glycerol
Saccharomyces cerevisiae
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
?
CTP + 1,2-diacyl-sn-glycerol
Saccharomyces cerevisiae BY4742
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
?
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.018
-
wild-type cells, pH 7.5, 30°C
Saccharomyces cerevisiae
130
-
DAG kinase in the membrane fraction of galactose-grown cells, pH 7.5, 30°C
Saccharomyces cerevisiae
Storage Stability (protein specific)
Storage Stability
Organism
-80°C, the enzyme preparation is completely stable for at least 3 months of storage at, and is stable to several cycles of freezing and thawing
Saccharomyces cerevisiae
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
CTP + 1,2-diacyl-sn-glycerol
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
687738
Saccharomyces cerevisiae
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
CTP + 1,2-diacyl-sn-glycerol
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity
687738
Saccharomyces cerevisiae BY4742
CDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
CTP + 1,2-dioleoyl-sn-glycerol
the CTP transferase domain is sufficient for diacylglycerol kinase activity
687738
Saccharomyces cerevisiae
CDP + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
-
-
?
CTP + 1,2-dioleoyl-sn-glycerol
the CTP transferase domain is sufficient for diacylglycerol kinase activity
687738
Saccharomyces cerevisiae BY4742
CDP + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
-
-
?
dCTP + 1,2-diacyl-sn-glycerol
both a substrate and competitive inhibitor
687738
Saccharomyces cerevisiae
dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
dCTP + 1,2-diacyl-sn-glycerol
both a substrate and competitive inhibitor
687738
Saccharomyces cerevisiae BY4742
dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
More
Dgk1p contains a CTP transferase domain, the CTP transferase domain is sufficient for diacylglycerol kinase activity
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
-
stable up to, labile above
Saccharomyces cerevisiae
70
-
loss of about 70% of the activity
Saccharomyces cerevisiae
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
7.5
-
Saccharomyces cerevisiae
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6.5
9
activity range, profile overview
Saccharomyces cerevisiae
General Information
General Information
Commentary
Organism
additional information
Dgk1p contains a CTP transferase domain that is present in the SEC59-encoded dolichol kinase and CDS1-encoded CDP-diacylglycerol synthase enzymes
Saccharomyces cerevisiae
physiological function
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity. DGK1 counteracts the PAH1-encoded PA phosphatase
Saccharomyces cerevisiae
General Information (protein specific)
General Information
Commentary
Organism
additional information
Dgk1p contains a CTP transferase domain that is present in the SEC59-encoded dolichol kinase and CDS1-encoded CDP-diacylglycerol synthase enzymes
Saccharomyces cerevisiae
physiological function
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity. DGK1 counteracts the PAH1-encoded PA phosphatase
Saccharomyces cerevisiae
Other publictions for EC 2.7.1.174
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)
738595
Qiu
Transcription factor Reb1p reg ...
Saccharomyces cerevisiae
J. Biol. Chem.
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29124-29133
2013
-
-
-
-
-
-
-
-
1
2
-
1
-
2
-
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
-
1
-
-
-
1
2
2
1
-
-
687738
Han
Characterization of the yeast ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4742
J. Biol. Chem.
283
20443-20453
2008
8
-
1
-
5
-
17
1
1
2
-
2
-
6
-
-
-
-
-
-
2
1
6
1
1
-
2
-
1
1
-
1
1
-
1
8
-
1
1
-
5
-
1
17
1
1
1
2
-
2
-
-
-
-
-
-
2
1
6
1
1
-
2
-
1
1
-
-
-
2
2
-
-
-
693092
Han
An unconventional diacylglycer ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae BY4742
J. Biol. Chem.
283
20433-20442
2008
-
-
1
-
1
-
-
-
2
1
-
4
-
4
-
-
-
-
-
-
-
-
6
-
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
-
1
-
-
-
-
-
2
1
-
4
-
-
-
-
-
-
-
-
6
-
1
-
-
-
1
-
-
-
-
4
4
-
-
-