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Literature summary for 2.7.1.107 extracted from

  • Jerga, A.; Miller, D.J.; White, S.W.; Rock, C.O.
    Molecular determinants for interfacial binding and conformational change in a soluble diacylglycerol kinase (2009), J. Biol. Chem., 284, 7246-7254.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
dioleoyl-phosphatidylglycerol
-
Staphylococcus aureus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Staphylococcus aureus

Protein Variants

Protein Variants Comment Organism
D124A mutant shows strongly reduced activity Staphylococcus aureus
D216A mutant shows strongly reduced activity Staphylococcus aureus
D271A mutant shows almost no activity Staphylococcus aureus
D68A mutant shows almost no activity Staphylococcus aureus
D97A mutant shows strongly reduced activity Staphylococcus aureus
E273A mutant shows almost no activity Staphylococcus aureus
K15A mutant shows reduced activity Staphylococcus aureus
K15A/K165A mutant shows strongly reduced activity Staphylococcus aureus
K165A mutant shows reduced activity Staphylococcus aureus
N96A mutant shows strongly reduced activity Staphylococcus aureus
R100A mutant shows reduced activity Staphylococcus aureus
R20A mutant shows wild type activity Staphylococcus aureus
T94A mutant shows strongly reduced activity Staphylococcus aureus

Localization

Localization Comment Organism GeneOntology No. Textmining
soluble
-
Staphylococcus aureus
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for activity, interfacial binding of DgkB requires a Mg2+-dependent conformational change Staphylococcus aureus

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus Q6GFF9
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Staphylococcus aureus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0004
-
mutant enzyme D271A, specific activity is less than 0.0004 micromol/min/mg, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0004
-
mutant enzyme D68A, specific activity is less than 0.0004 micromol/min/mg, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0004
-
mutant enzyme E273A, specific activity is less than 0.0004 micromol/min/mg, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0023
-
mutant enzyme N96A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0024
-
mutant enzyme D124A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0028
-
mutant enzyme K15A/K165A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0031
-
mutant enzyme T94A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0036
-
mutant enzyme D97A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0062
-
mutant enzyme D216A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0122
-
mutant enzyme K15A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0139
-
mutant enzyme K165A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0168
-
mutant enzyme R100A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0249
-
wild type enzyme, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus
0.0256
-
mutant enzyme R20A, in 50 mM HEPES, pH 7.4, 150 mM LiCl, 10 mM MgCl2, 5 mM ATP, and lipid vesicles Staphylococcus aureus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 1,2-diacylglycerol
-
Staphylococcus aureus ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
?
additional information 1,2-diacylglycerol embedded in unilamellar dioleoyl-phosphatidylcholine vesicles is not a substrate for DgkB Staphylococcus aureus ?
-
?

Synonyms

Synonyms Comment Organism
DAG kinase
-
Staphylococcus aureus
DgkB
-
Staphylococcus aureus

Cofactor

Cofactor Comment Organism Structure
ATP
-
Staphylococcus aureus