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Information on EC 2.7.1.107 - diacylglycerol kinase (ATP) and Organism(s) Escherichia coli and UniProt Accession P0ABN1
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2.7.1.107 diacylglycerol kinase (ATP)IUBMB Comments
Involved in synthesis of membrane phospholipids and the neutral lipid triacylglycerol. Activity is stimulated by certain phospholipids [4,7]. In plants and animals the product 1,2-diacyl-sn-glycerol 3-phosphate is an important second messenger. cf. EC 2.7.1.174, diacylglycerol kinase (CTP).
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Word Map
- 2.7.1.107
-
phosphatidic
- phospholipase
- phospholipid
- pkc
- phosphatidylinositol
-
phorbol
- isozymes
- phosphoinositide
- phosphatidylcholine
- inositol
- ceramide
- lipase
-
arachidonic
- deoxyguanosine
- deoxycytidine
-
ef-hands
- phosphatidylserine
- 13-acetate
-
polyphosphoinositide
-
phosphohydrolase
-
prelabeled
- deoxynucleoside
- staurosporine
- sphingosine
- pip2
-
pleckstrin
- 4,5-bisphosphate
-
1,4,5-trisphosphate
- 4-phosphate
- deoxyribonucleoside
- analysis
- marcks
-
kinase-dead
- medicine
- phosphatidylethanol
-
transphosphatidylation
- dioctanoylglycerol
- hypospadias
- ptdoh
- 5-kinase
- agriculture
-
3hglycerol
- monoacylglycerols
-
anergy
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
dgk, diacylglycerol kinase, dag kinase, dg kinase, dgkzeta, dagk, dgkalpha, dgk-zeta, diglyceride kinase, dgkepsilon, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
1,2-diacylglycerol kinase
-
-
-
-
adenosine 5'-triphosphate:1,2-diacylglycerol 3-phosphotransferase
-
-
-
-
arachidonoyl-specific diacylglycerol kinase
-
-
-
-
ATP:diacylglycerol phosphotransferase
-
-
-
-
DAGK
DAGKalpha
-
-
-
-
DG kinase
-
-
-
-
DGK
DGK-alpha
-
-
-
-
DGK-theta
-
-
-
-
DGKbeta
-
-
-
-
DGKdelta
-
-
-
-
DGKgamma
-
-
-
-
DGKiota
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-
-
-
DGKksi
-
-
-
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diacylglycerol kinase
-
-
-
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diacylglycerol kinase (ATP dependent)
-
-
-
-
diacylglycerol:ATP kinase
-
-
-
-
diglyceride kinase
-
-
-
-
kinase (phosphorylating), 1,2-diacylglycerol
-
-
-
-
kinase, 1,2-diacylglycerol (phosphorylating)
-
-
-
-
sn-1,2-diacylglycerol kinase
-
-
-
-
DAGK
-
-
-
-
DGK
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + 1,2-diacyl-sn-glycerol = ADP + 1,2-diacyl-sn-glycerol 3-phosphate
random equilibrium mechanism
-
ATP + 1,2-diacyl-sn-glycerol = ADP + 1,2-diacyl-sn-glycerol 3-phosphate
analysis of the catalytic kinase mechanism molecular dynamics and quantum mechanics calculations, and density functional theory calculations, overview. The active site is relatively open and able to accommodate ligands in multiple orientations, suggesting that the optimization of binding orientations and conformational changes occurs prior to actual phosphoryl transfer
ATP + 1,2-diacyl-sn-glycerol = ADP + 1,2-diacyl-sn-glycerol 3-phosphate
analysis of the catalytic mechanism using crystal structure anaysis, structure-function analysis, mutagenesis studies, molecular dynamics simulations, and density functional theory modelling, overview
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
phospho group transfer
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-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
ATP:1,2-diacyl-sn-glycerol 3-phosphotransferase
Involved in synthesis of membrane phospholipids and the neutral lipid triacylglycerol. Activity is stimulated by certain phospholipids [4,7]. In plants and animals the product 1,2-diacyl-sn-glycerol 3-phosphate is an important second messenger. cf. EC 2.7.1.174, diacylglycerol kinase (CTP).
CAS REGISTRY NUMBER
COMMENTARY
60382-71-0
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + 1,2-diacylglycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
ATP + 1,2-dioleoylglycerol
ADP + 1,2-dioleoylglycerol 3-phospate
modeling of lipid substrate binding, involving residues Arg9, Ser17, Ser98 and Glu69, overview
-
-
?
1,2-dipalmitoyl-sn-glycerol + GTP
GDP + 1,2-dipalmitoyl-sn-glycerol 3-phosphate
-
-
-
-
?
2'-deoxy-ATP + sn-1,2-dihexanoylglycerol
2'-deoxy-ADP + sn-1,2-dihexanoylglycerol 3-phosphate
-
-
-
-
?
ADP + sn-1,2-dihexanoylglycerol
AMP + sn-1,2-dihexanoylglycerol 3-phosphate
-
MgADP- is a very poor phosphoryl donor
-
-
?
ATP + 1,2-diacylglycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
i.e. phosphatidic acid
-
r
ATP + 1,2-dihexanoylglycerol
ADP + 1,2-dihexanoylglycerol 3-phosphate
-
-
-
-
?
ATP + sn-1,2-dihexanoylglycerol
ADP + sn-1,2-dihexanoylglycerol 3-phosphate
-
-
-
-
?
ATP + sn-1,2-dioctanoylglycerol
ADP + sn-1,2-dioctanoylglycerol 3-phosphate
-
-
-
-
?
ATP + sn-1,2-dioleoylglycerol
ADP + sn-1,2-dioleoylglycerol 3-phosphate
-
-
-
-
?
GTP + sn-1,2-dihexanoylglycerol
GDP + sn-1,2-dihexanoylglycerol 3-phosphate
-
-
-
-
?
ITP + sn-1,2-dihexanoylglycerol
IDP + sn-1,2-dihexanoylglycerol 3-phosphate
-
-
-
-
?
ATP + 1,2-diacyl-sn-glycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
ATP + 1,2-diacyl-sn-glycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
enzyme DgkA primarily recognizes diacylglycerol in the glycerol backbone and ester linkages but not the fatty acyl group
-
-
?
ATP + 1,2-diacyl-sn-glycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
-
?
ATP + 1,2-diacyl-sn-glycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
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the enzyme functions to recycle diacylglycerol which is generated largely as a by-product of membrane-derived oligosaccharide biosynthesis
-
-
?
additional information
?
-
DgkA also has ATPase activity which is about 25% of its kinase activity
-
-
?
additional information
?
-
-
sn-1,3-dioleoylglycerol is not a substrate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + 1,2-diacyl-sn-glycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
ATP + 1,2-diacylglycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
-
-
?
ATP + 1,2-diacyl-sn-glycerol
ADP + 1,2-diacyl-sn-glycerol 3-phosphate
-
the enzyme functions to recycle diacylglycerol which is generated largely as a by-product of membrane-derived oligosaccharide biosynthesis
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Cd2+
-
enzyme requires a free divalent metal cation: Mg2+, Mn2+, Co2+, Cd2+ or Zn2+
Co2+
-
enzyme requires a free divalent metal cation: Mg2+, Mn2+, Co2+, Cd2+ or Zn2+
Mg2+
Mn2+
-
enzyme requires a free divalent metal cation: Mg2+, Mn2+, Co2+, Cd2+ or Zn2+. Ka for Mg2+ is 3.4 mM
Zn2+
-
enzyme requires a free divalent metal cation: Mg2+, Mn2+, Co2+, Cd2+ or Zn2+
Mg2+
-
enzyme requires a free divalent metal cation: Mg2+, Mn2+, Co2+, Cd2+ or Zn2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
diacylglycerol 3-phosphate
-
the enzyme apoprotein is attributed to a novel feedback activation involving diacylglycerol 3-phosphate
Lipid
-
purified enzyme is completely inactive unless a lipid is added to the assay buffer containing Triton X-100
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
Km for dioleoylglycerol is 0.92 mol%
-
additional information
additional information
-
kinetics, recombinant wild-type and mutant enzymes
-
additional information
additional information
-
Km-value in mol% for wild-type 9.2, mutant C46A/C113A 4.9, mutant C46A/C113A/E28C 5.7, mutant C46A/C113A/A29C 7.4, mutant C46A/C113AA30C 27.2, mutant C46A/C113A/F31C 17.2, mutant C46A/C113A/R32C 12.9, mutant C46A/C113A/Q33C 12.1, mutant C46A/C113A/E34C 13.7
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
Ki-value for adenosine 5'-tetraphosphoryl-3-O-(1,2-dihexanoyl)-sn-glycerol is 0.036 mol%
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
the enzyme is an integral membrane protein, a trimer situated with half its bulk in the membrane and half in the cytosol
-
the region of transmembrane helix 1 spanning the hydrophobic core of the bilayer runs from Glu28 on the cytoplasmic side to Asp49 or Val50 on the periplasmic side. This locates the charged/polar cluster 32RQE34 within the hydrophobic core of the bilayer. Hydrophobic matching between the protein and the surrounding lipid bilayer is highly efficient
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
physiological function
diacylglycerol kinase catalyzes the ATP-dependent phosphorylation of diacylglycerol to phosphatidic acid for use in shuttling water-soluble components to membrane-derived oligosaccharide and lipopolysaccharide in the cell envelope of Gram-negative bacteria
additional information
evolution
as the smallest kinase known, it shares no sequence homology with conventional kinases and possesses a distinct trimer structure. The phosphorylation reaction of diacylglycerol kinase features the same phosphoryl transfer mechanism as other kinases, despite its unique structural properties. DgkA appears to be an evolutionarily optimized enzyme and its chemical reaction rate approaches the substrate diffusion-controlled rate limit
evolution
DgkA is a unique kinase with a distinctive active site. It has no recognizable nucleotide sequence or structural binding motifs
additional information
1,2-dioctanoylglycerol is first docked into the active site of the crystal structure of DgkA, PDB ID 3ZE5, followed by construction of a ternary complex model by docking co-factor ATP and substrate 1,2-dioctanoylglycerol into the active site of DgkA. The complex of DgkA is optimized and equilibrated by molecular dynamics simulation in the lipid bilayer. The phosphotransfer reaction catalyzed by DgkA is then investigated through the hybrid density functional theory method B3LYP. Important role of the surface helix in the active site formation
additional information
Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76 both of which are essential, the gamma-phosphate of ATP is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane, catalytic mechanism, overview. The putative catalytic site resides on the protein at the membrane/cytosol interface where the reactive moieties of the two substrates, with disparate polarities, come together for reaction. The ternary complex site, asBC, contains zinc-ACP and two lipid substrates. The gaamma-phosphate of the ATP analogue is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane
additional information
-
Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76 both of which are essential, the gamma-phosphate of ATP is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane, catalytic mechanism, overview. The putative catalytic site resides on the protein at the membrane/cytosol interface where the reactive moieties of the two substrates, with disparate polarities, come together for reaction. The ternary complex site, asBC, contains zinc-ACP and two lipid substrates. The gaamma-phosphate of the ATP analogue is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane
additional information
DgkA catalyzes phosphoryl transfer expected to take place at a polar/apolar interface
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homotrimer
trimer
monomer
homotrimer
a homotrimeric enzyme with three transmembrane helices and an N-terminal amphiphilic helix per monomer. Bound lipid substrate and docked ATP identify the putative active site which is of the composite, shared site type. Each subunit within a trimer forms a bundle of 3 transmembrane helices. The three subunits are arranged around an approximate 3-fold symmetry axis that passes through the center of the trimer normal to the membrane plane, enzyme structure analysis, overview
trimer
optimized structures of the reactant, transition state and product with Mg2+ coordination, overview
trimer
the enzyme crafts three catalytic and substrate-binding sites centred about the membrane/cytosol interface
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified kinase captured as a ternary complex with bound lipid substrate and an ATP analogue, method optimization, microseeding, the precipitant solution contains 0.2% v/v MPD, 0.1 M NaCl, 0.05 M Na3C6H5O7, pH 5.6, 20°C, X-ray diffraction structure determination and analysis at resolutions of 2.18-3.20 A
purified recombinanz wild-type and DELTA4 and DELTA7 mutant enzymes in complex with lipid substrate and ATP, 20°C, X-ray diffraction structure determination and analysis at 2.05 A resolution, structure mmodeling. The kinase adopts a functional form in the crystal. Domain swapping, a key feature of the solution form, is not observed in the crystal structures. Crystals of mutant DELTA7 DgkA are soaked with the ATP analogue, adenylylmethylenediphosphate (Mg2+-AMPPCP). This causes the crystals to dissolve. Additional soaking with ATP, ADP, AMP, ATP?S, AMPPNP and dATP but not with GTP, CTP, UTP or TTP leads to crystal dissolution
sitting-drop vapour-diffusion method. Crystals belong to space group P2(1), with unit-cell parameters a = 42.4, b = 166.1, c = 48.5 A, beta = 96.97°
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A13K
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
A13R
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
A30L
site-directed mutagenesis, the mutant shows 93% reduced activity compared to the wild-type enzyme
D80A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
D80E
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D80N
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D81A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D81K
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D95A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
D95E
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D95N
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
E28A
site-directed mutagenesis, the mutation principally affects the binding of the Zn2+ ion, In the absence of the E28 side chain the zinc ions become purely coordinated by E76 and the ATP phosphates, the mutant shows highly reduced activity compared to the wild-type enzyme
E28D
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
E28N
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
E28Q
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
E28R
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
E34A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E34D
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E34Q
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E69A
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
E69D
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
E69Q
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
E76A
E76D
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
E76Q
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
G20A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
K94A
K94 coordinates both alpha-phosphate and N7 of the adenine ring of ATP, the loss of the basic side-chain releases the adenine of ATP and the binding is lost, almost inactive mutant
K94M
K94 coordinates both alpha-phosphate and N7 of the adenine ring of ATP, the loss of the basic side-chain releases the adenine of ATP and the binding is lost, the mutant shows highly reduced activity compared to the wild-type enzyme
K94R
K94 coordinates both alpha-phosphate and N7 of the adenine ring of ATP, the loss of the basic side-chain releases the adenine of ATP and the binding is lost, almost inactive mutant
N72A
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
N72D
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
N72Q
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R32A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R32K
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
R9A
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R9E
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R9H
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
R9K
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme
S17A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S73A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S98A
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
Y86A
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
Y86F
site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme
A14Q
-
significantly impaired catalytic function, without evidence of gross structural alterations, subunit mixing experiments of mutant enzymes, subunit mixing experiments of mutant enzymes
C46A/C113A
-
mutant lacking all Cys residues. Activity is slightly higher than wild-type
C46A/C113A/A29C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 64% trimer formation compared to wild-type
C46A/C113A/A30C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 79% trimer formation compared to wild-type
C46A/C113A/E28C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 93% trimer formation compared to wild-type
C46A/C113A/Q33C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 77% trimer formation compared to wild-type
C46A/C113A/R32C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 63% trimer formation compared to wild-type
C46A/C113AE34C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 100% trimer formation compared to wild-type
C46A/C113AF31C
-
introduction of Cys residue at transmembrane helix 1 into mutant lacking the native Cys residues. Low activity mutant, 72% trimer formation compared to wild-type
D95N
-
significantly impaired catalytic function, without evidence of gross structural alterations. Km-value for MgATP2- raises 18fold, subunit mixing experiments of mutant enzymes
E76L
-
significantly impaired catalytic function, without evidence of gross structural alterations, subunit mixing experiments of mutant enzymes
I110P
-
mutant enzyme can not be purified because its expression is toxic to the Escherichia coli host
I110R
-
mutant enzyme can not be purified because its expression is toxic to the Escherichia coli host
I110W
-
mutant is highly misfolding while at the same time being more stable than the wild-type protein
I110Y
-
mutant exhibits enhanced stability but folds with an efficiency similar to that of the wild type
K94L
-
significantly impaired catalytic function, without evidence of gross structural alterations. Km-value for MgATP2- raises 13fold, subunit mixing experiments of mutant enzymes
N72S
-
significantly impaired catalytic function, without evidence of gross structural alterations, subunit mixing experiments of mutant enzymes
W18L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
W18L/W25L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
W18L/W25L/W47L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
W18L/W25L/W47L/W117L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
W18L/W47L/W117L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
W25L
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
additional information
construction of a thermostabilized DELTA4 (4 changes relative to wild-type) form of DgkA using the DELTA7 structure, overview
E76A
site-directed mutagenesis, inactive mutant, Asn72 plays a key role in catalysis. Its side-chain amide bridges Glu69 and Glu76
E76A
site-directed mutagenesis, the mutation principally affects the binding of the Zn2+ ion
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
100
additional information
-
addition of phospholipids provides some protection from thermal inactivation
100
-
the membrane preparation shows a threefold stimulation by heating for 5 min followed by a gradual loss of activity, half-life: about 1 h. Butane-1-ol-dissolved enzyme has a half-life of about 45 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
lipid activators stabilize the enzyme agisnt inactivation induced by diacylglycerol. Mg2+ and Mn2+ show only a small stabilization effect both in presence and in absence of 10 mol% phosphatidylglycerol
-
when cosolubilized with diacylglycerol in octylglucoside micelles, the enzyme undergoes rapid irreversible inactivation
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene dgkA, recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain WH1061 by nickel affinity chromatography and gel filtration. The size-exclusion chromatography step has no effect on specific activity
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene dgkA, recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain WH1061
a 100fold overproduction is obtained when dgkA is placed on a hybrid plasmid under control of the lambdapl promoter
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Walsh, J.P.; Bell, R.M.
Diacylglycerol kinase from Escherichia coli
Methods Enzymol.
209
153-162
1992
Escherichia coli
Walsh, J.P.; Bell, R.M.
sn-1,2-Diacylglycerol kinase of Escherichia coli. Structural and kinetic analysis of the lipid cofactor dependence
J. Biol. Chem.
261
15062-15069
1986
Escherichia coli
Schaap, D.; de Widt, J.; van der Wal, J.; Vandekerckhove, J.; van Damme, J.; Gussow, D.; Ploegh, H.L.; van Blitterswijk, W.J.; van der Bend, R.L.
Purification, cDNA-cloning and expression of human diacylglycerol kinase
FEBS Lett.
275
151-158
1990
Escherichia coli, Homo sapiens
Russ, E.; Kaiser, U.; Sandermann, H.
Lipid-dependent membrane enzymes. Purification to homogeneity and further characterization of diacylglycerol kinase from Escherichia coli
Eur. J. Biochem.
171
335-342
1988
Escherichia coli
Bohnenberger, E.; Sandermann, H.
Lipid dependence of diacylglycerol kinase from Escherichia coli
Eur. J. Biochem.
132
645-650
1983
Escherichia coli
Bohnenberger, E.; Sandermann, H.
Diglyceride kinase from Escherichia coli. Purification in organic solvent and some properties of the enzyme
Eur. J. Biochem.
94
401-407
1979
Escherichia coli
Loomis, C.R.; Walsh, J.P.; Bell, R.M.
sn-1,2-Diacylglycerol kinase of Escherichia coli. Purification, reconstitution, and partial amino- and carboxyl-terminal analysis
J. Biol. Chem.
260
4091-4097
1985
Escherichia coli
Lau, F.W.; Chen, X.; Bowie, J.U.
Active sites of diacylglycerol kinase from Escherichia coli are shared between subunits
Biochemistry
38
5521-5527
1999
Escherichia coli
Badola, P.; Sanders, C.R.2nd.
Escherichia coli diacylglycerol kinase is an evolutionarily optimized membrane enzyme and catalyzes direct phosphoryl transfer
J. Biol. Chem.
272
24176-24182
1997
Escherichia coli
Clark, E.H.; East, J.M.; Lee, A.G.
The role of tryptophan residues in an integral membrane protein: diacylglycerol kinase
Biochemistry
42
11065-11073
2003
Escherichia coli
Bakali, M.A.; Nordlund, P.; Hallberg, B.M.
Expression, purification, crystallization and preliminary diffraction studies of the mammalian DAG kinase homologue YegS from Escherichia coli
Acta Crystallogr. Sect. F
62
295-297
2006
Escherichia coli
Mi, D.; Kim, H.J.; Hadziselimovic, A.; Sanders, C.R.
Irreversible misfolding of diacylglycerol kinase is independent of aggregation and occurs prior to trimerization and membrane association
Biochemistry
45
10072-10084
2006
Escherichia coli
Jittikoon, J.; East, J.M.; Lee, A.G.
A fluorescence method to define transmembrane alpha-helices in membrane proteins: studies with bacterial diacylglycerol kinase
Biochemistry
46
10950-10959
2007
Escherichia coli
Van Horn, W.; Kim, H.; Ellis, C.; Hadziselimovic, A.; Sulistijo, E.; Karra, M.; Tian, C.; Soennichsen, F.; Sanders, C.
Solution nuclear magnetic resonance structure of membrane-integral diacylglycerol kinase
Science
324
1726-1729
2009
Escherichia coli (P0ABN1), Escherichia coli
Li, D.; Stansfeld, P.J.; Sansom, M.S.; Keogh, A.; Vogeley, L.; Howe, N.; Lyons, J.A.; Aragao, D.; Fromme, P.; Fromme, R.; Basu, S.; Grotjohann, I.; Kupitz, C.; Rendek, K.; Weierstall, U.; Zatsepin, N.A.; Cherezov, V.; Liu, W.; Bandaru, S.; English, N.J.; Gati, C.; Barty, A.; Yefanov, O.; Chapman, H.N.; Diederichs, K.; Messerschmidt, M.; Boutet, S.; Williams, G.J.; Seibert, M.M.; Caffrey, M.
Ternary structure reveals mechanism of a membrane diacylglycerol kinase
Nat. Commun.
6
10140
2015
Escherichia coli (P0ABN1), Escherichia coli
Li, D.; Lyons, J.A.; Pye, V.E.; Vogeley, L.; Aragao, D.; Kenyon, C.P.; Shah, S.T.; Doherty, C.; Aherne, M.; Caffrey, M.
Crystal structure of the integral membrane diacylglycerol kinase
Nature
497
521-524
2013
Escherichia coli (P0ABN1)
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