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Literature summary extracted from

  • Han, G.S.; OHara, L.; Siniossoglou, S.; Carman, G.M.
    Characterization of the yeast DGK1-encoded CTP-dependent diacylglycerol kinase (2008), J. Biol. Chem., 283, 20443-20453.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment 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 Cloned (Comment) 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

Protein Variants

EC Number Protein Variants Comment 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 Comment 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 Comment 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 Comment Organism GeneOntology No. Textmining
2.7.1.174 membrane
-
Saccharomyces cerevisiae 16020
-

Metals/Ions

EC Number Metals/Ions Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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 UniProt Comment 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] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.174 CTP + 1,2-diacyl-sn-glycerol in vivo functions of Dgk1p are specifically due to its diacylglycerol kinase activity 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 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 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 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 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 Saccharomyces cerevisiae BY4742 dCDP + 1,2-diacyl-sn-glycerol 3-phosphate
-
?

Subunits

EC Number Subunits Comment Organism
2.7.1.174 More Dgk1p contains a CTP transferase domain, the CTP transferase domain is sufficient for diacylglycerol kinase activity Saccharomyces cerevisiae

Synonyms

EC Number Synonyms Comment Organism
2.7.1.108 Sec59p
-
Saccharomyces cerevisiae
2.7.1.174 CTP-dependent diacylglycerol kinase
-
Saccharomyces cerevisiae
2.7.1.174 DGK1
-
Saccharomyces cerevisiae
2.7.1.174 Dgk1p
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.174 30
-
assay at Saccharomyces cerevisiae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment 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 Comment Organism
2.7.1.174 7 7.5
-
Saccharomyces cerevisiae

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.1.174 6.5 9 activity range, profile overview Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment 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 Comment 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 Comment Organism Inhibitor Structure
2.7.1.174 0.1
-
pH 7.5, 30°C Saccharomyces cerevisiae N-ethylmaleimide

General Information

EC Number General Information Comment 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