BRENDA - Enzyme Database

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
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.174
dolichol
-
Saccharomyces cerevisiae
2.7.1.174
additional information
diacylglycerol kinase activity is stimulated by major membrane phospholipids, overview
Saccharomyces cerevisiae
2.7.1.174
phosphatidate
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylcholine
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylethanolamine
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylglycerol
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylinositol
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylserine
-
Saccharomyces cerevisiae
Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.174
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
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.174
D177A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
2.7.1.174
G184A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
2.7.1.174
K77A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing an almost complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
2.7.1.174
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
2.7.1.174
R76A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.174
cardiolipin
-
Saccharomyces cerevisiae
2.7.1.174
CDP-diacylglycerol
-
Saccharomyces cerevisiae
2.7.1.174
ceramide
-
Saccharomyces cerevisiae
2.7.1.174
dCTP
both a substrate and competitive inhibitor
Saccharomyces cerevisiae
2.7.1.174
diacylglycerol diphosphate
-
Saccharomyces cerevisiae
2.7.1.174
lyso-phosphatidate
-
Saccharomyces cerevisiae
2.7.1.174
Mn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
2.7.1.174
additional information
the enzyme is inhibited by sphingoid bases
Saccharomyces cerevisiae
2.7.1.174
N-ethylmaleimide
-
Saccharomyces cerevisiae
2.7.1.174
phytosphingosine
-
Saccharomyces cerevisiae
2.7.1.174
R59022
25% inhibition at 0.05 M
Saccharomyces cerevisiae
2.7.1.174
R59949
15% inhibition at 0.05 M
Saccharomyces cerevisiae
2.7.1.174
sphinganine
-
Saccharomyces cerevisiae
2.7.1.174
sphingosine
-
Saccharomyces cerevisiae
2.7.1.174
triacylglycerol
-
Saccharomyces cerevisiae
2.7.1.174
Triton X-100
inhibition kinetics, overview. No inhibition by monoacylglycerol
Saccharomyces cerevisiae
2.7.1.174
Zn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.174
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
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.1.174
membrane
-
Saccharomyces cerevisiae
16020
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.174
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
2.7.1.174
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)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.174
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
-
-
?
2.7.1.174
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.108
Saccharomyces cerevisiae
-
CTP transferase domain that is also present in CDS1-encoded CDP-diacylglycerolsynthase
-
2.7.1.174
Saccharomyces cerevisiae
Q12382
gene DGK1 or YOR311C
-
2.7.1.174
Saccharomyces cerevisiae BY4742
Q12382
gene DGK1 or YOR311C
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.174
0.018
-
wild-type cells, pH 7.5, 30°C
Saccharomyces cerevisiae
2.7.1.174
130
-
DAG kinase in the membrane fraction of galactose-grown cells, pH 7.5, 30°C
Saccharomyces cerevisiae
Storage Stability
EC Number
Storage Stability
Organism
2.7.1.174
-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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.174
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
-
-
-
?
2.7.1.174
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
-
-
-
?
2.7.1.174
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
-
-
-
?
2.7.1.174
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
-
-
-
?
2.7.1.174
dCTP + 1,2-diacyl-sn-glycerol
both a substrate and competitive inhibitor
687738
Saccharomyces cerevisiae
dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
2.7.1.174
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
EC Number
Subunits
Commentary
Organism
2.7.1.174
More
Dgk1p contains a CTP transferase domain, the CTP transferase domain is sufficient for diacylglycerol kinase activity
Saccharomyces cerevisiae
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.174
30
-
assay at
Saccharomyces cerevisiae
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.1.174
40
-
stable up to, labile above
Saccharomyces cerevisiae
2.7.1.174
70
-
loss of about 70% of the activity
Saccharomyces cerevisiae
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.174
7
7.5
-
Saccharomyces cerevisiae
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.7.1.174
6.5
9
activity range, profile overview
Saccharomyces cerevisiae
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.108
CTP
-
Saccharomyces cerevisiae
2.7.1.174
CTP
dependent on
Saccharomyces cerevisiae
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.7.1.174
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
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
2.7.1.174
0.1
-
pH 7.5, 30°C
Saccharomyces cerevisiae
N-ethylmaleimide
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.174
dolichol
-
Saccharomyces cerevisiae
2.7.1.174
additional information
diacylglycerol kinase activity is stimulated by major membrane phospholipids, overview
Saccharomyces cerevisiae
2.7.1.174
phosphatidate
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylcholine
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylethanolamine
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylglycerol
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylinositol
-
Saccharomyces cerevisiae
2.7.1.174
phosphatidylserine
-
Saccharomyces cerevisiae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.174
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)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.108
CTP
-
Saccharomyces cerevisiae
2.7.1.174
CTP
dependent on
Saccharomyces cerevisiae
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.174
D177A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
2.7.1.174
G184A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing a loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
2.7.1.174
K77A
site-directed mutagenesis of a conserved residue within the CTP transferase domain causing an almost complete loss of diacylglycerol kinase activity
Saccharomyces cerevisiae
2.7.1.174
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
2.7.1.174
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)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
2.7.1.174
0.1
-
pH 7.5, 30°C
Saccharomyces cerevisiae
N-ethylmaleimide
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.174
cardiolipin
-
Saccharomyces cerevisiae
2.7.1.174
CDP-diacylglycerol
-
Saccharomyces cerevisiae
2.7.1.174
ceramide
-
Saccharomyces cerevisiae
2.7.1.174
dCTP
both a substrate and competitive inhibitor
Saccharomyces cerevisiae
2.7.1.174
diacylglycerol diphosphate
-
Saccharomyces cerevisiae
2.7.1.174
lyso-phosphatidate
-
Saccharomyces cerevisiae
2.7.1.174
Mn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
2.7.1.174
additional information
the enzyme is inhibited by sphingoid bases
Saccharomyces cerevisiae
2.7.1.174
N-ethylmaleimide
-
Saccharomyces cerevisiae
2.7.1.174
phytosphingosine
-
Saccharomyces cerevisiae
2.7.1.174
R59022
25% inhibition at 0.05 M
Saccharomyces cerevisiae
2.7.1.174
R59949
15% inhibition at 0.05 M
Saccharomyces cerevisiae
2.7.1.174
sphinganine
-
Saccharomyces cerevisiae
2.7.1.174
sphingosine
-
Saccharomyces cerevisiae
2.7.1.174
triacylglycerol
-
Saccharomyces cerevisiae
2.7.1.174
Triton X-100
inhibition kinetics, overview. No inhibition by monoacylglycerol
Saccharomyces cerevisiae
2.7.1.174
Zn2+
abolishes detectable DAG kinase activity
Saccharomyces cerevisiae
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
2.7.1.174
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)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.174
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)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.1.174
membrane
-
Saccharomyces cerevisiae
16020
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.174
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
2.7.1.174
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)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.174
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
-
-
?
2.7.1.174
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)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.174
0.018
-
wild-type cells, pH 7.5, 30°C
Saccharomyces cerevisiae
2.7.1.174
130
-
DAG kinase in the membrane fraction of galactose-grown cells, pH 7.5, 30°C
Saccharomyces cerevisiae
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
2.7.1.174
-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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.174
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
-
-
-
?
2.7.1.174
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
-
-
-
?
2.7.1.174
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
-
-
-
?
2.7.1.174
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
-
-
-
?
2.7.1.174
dCTP + 1,2-diacyl-sn-glycerol
both a substrate and competitive inhibitor
687738
Saccharomyces cerevisiae
dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
2.7.1.174
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)
EC Number
Subunits
Commentary
Organism
2.7.1.174
More
Dgk1p contains a CTP transferase domain, the CTP transferase domain is sufficient for diacylglycerol kinase activity
Saccharomyces cerevisiae
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.174
30
-
assay at
Saccharomyces cerevisiae
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.1.174
40
-
stable up to, labile above
Saccharomyces cerevisiae
2.7.1.174
70
-
loss of about 70% of the activity
Saccharomyces cerevisiae
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.174
7
7.5
-
Saccharomyces cerevisiae
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.7.1.174
6.5
9
activity range, profile overview
Saccharomyces cerevisiae
General Information
EC Number
General Information
Commentary
Organism
2.7.1.174
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
2.7.1.174
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)
EC Number
General Information
Commentary
Organism
2.7.1.174
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
2.7.1.174
physiological function
in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity. DGK1 counteracts the PAH1-encoded PA phosphatase
Saccharomyces cerevisiae