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show all sequences of 2.7.1.130

Mechanistic characterization of the tetraacyldisaccharide-1-phosphate 4-kinase LpxK involved in lipid A biosynthesis

Emptage, R.P.; Pemble, C.W.; York, J.D.; Raetz, C.R.; Zhou, P.; Biochemistry 52, 2280-2290 (2013)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C41(DE3) membranes
Aquifex aeolicus
Crystallization (Commentary)
Crystallization
Organism
purified LpxK in complex with the ATP analogue AMPPCP in the closed catalytically competent conformation, sitting drop vapor diffusion method, mixing of 0.70 ml well solution with 0.01 ml sample solution, containing four parts of a reservoir solution consisting of 50% v/v MPD and 0.1 M HEPES, pH 7.5, and one part protein solution containing 13 mg/ml enzyme LpxK, 4.3 mM AMP-PCP, 1 mM EDTA, 0.5% w/v DDM, 540 mM NaCl, 14% v/v glycerol, and 35 mM HEPES, pH 8.0, 20°C, 1 month, or by microseeding, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement using ADP-Mg2+ LpxK structure, PDB ID 4EHY, as the search model with all ligands removed, and AMP-PCP is subsequently added to the model. Purified LpxK in complex with ATP in a pre-catalytic binding state, mixing of seventeen parts of a reservoir solution consisting of 60% v/v MPD and 0.1 M HEPES, pH 7.5, and three parts protein solution containing 7.4 mg/ml enzyme LpxK, 10 mM ATP, 1 mM EDTA, 0.35% w/v DDM, 700 mM NaCl, 18.5% v/v glycerol, and 45 mM HEPES, pH 8.0, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using LpxK structure, PDB ID 4EHX, as the search model, ATP is subsequently added to the model. Purified LpxK in complex with a chloride anion in an inhibitory conformation of the nucleotide-binding P-loop, mixing of three parts of a reservoir solution consisting of 40% v/v MPD and 0.1 M HEPES, pH 7.5, and one part protein solution containing 8.3 mg/ml LpxK, 4 mM methyl 2-acetamido-2-deoxy-beta-D-glucopyranoside, 0.35% w/v DDM, 625 mM NaCl, 17% v/v glycerol, and 45 mM HEPES, pH 8.0, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using the apo enzyme LpxK structure, PDB ID 4EHX, as search model and spherical active site density refines well as a chloride ion
Aquifex aeolicus
Engineering
Amino acid exchange
Commentary
Organism
D138A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D138N
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D139A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D139N
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D260A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D99A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D99E
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D99N
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E100A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E100D
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E100Q
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
H261A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
K51A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
S49A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
S53A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
T52A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
Y74A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
Inhibitors
Inhibitors
Commentary
Organism
Structure
Ca2+
-
Aquifex aeolicus
Cu2+
-
Aquifex aeolicus
Fe3+
-
Aquifex aeolicus
Ni2+
-
Aquifex aeolicus
Zn2+
-
Aquifex aeolicus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
bi-substrate kinetics, steady-state kinetics
Aquifex aeolicus
0.0011
-
ATP
pH 8.5, 30°C, recombinant mutant D99N
Aquifex aeolicus
0.0012
-
ATP
pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
0.0013
-
ATP
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0014
-
ATP
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.0015
-
ATP
pH 8.5, 30°C, recombinant mutant K51A
Aquifex aeolicus
0.0016
-
ATP
pH 8.5, 30°C, recombinant mutant D260A
Aquifex aeolicus
0.0017
-
ATP
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.0019
-
ATP
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.002
-
ATP
pH 8.5, 30°C, recombinant mutant E100D
Aquifex aeolicus
0.0022
-
ATP
pH 8.5, 30°C, recombinant mutant T52A
Aquifex aeolicus
0.0023
-
ATP
pH 8.5, 30°C, recombinant mutant D99E
Aquifex aeolicus
0.0027
-
ATP
pH 8.5, 30°C, recombinant mutant S49A
Aquifex aeolicus
0.0028
-
ATP
pH 8.5, 30°C, recombinant mutant D99A
Aquifex aeolicus
0.0029
-
ATP
pH 8.5, 30°C, recombinant mutant E100Q
Aquifex aeolicus
0.0032
-
ATP
pH 8.5, 30°C, recombinant mutant D138N
Aquifex aeolicus
0.004
-
ATP
pH 8.5, 30°C, recombinant mutant D138A
Aquifex aeolicus
0.005
-
ATP
pH 8.5, 30°C, recombinant mutant D139A
Aquifex aeolicus
0.006
-
ATP
pH 8.5, 30°C, recombinant mutant S53A
Aquifex aeolicus
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
inner membrane
cytosol-facing, membrane binding through a hydrophobic lower face assisted by surrounding basic residues of the N-terminal core domain
Aquifex aeolicus
-
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Cl-
structural basis for anion inhibition of LpxK, modeling, overview
Aquifex aeolicus
Co2+
activates
Aquifex aeolicus
Mg2+
required, activates, optimal activity is observed at an equimolar ratio of ATP to Mg2+, with a decrease in activity at higher amounts of the divalent cation
Aquifex aeolicus
Mn2+
activates
Aquifex aeolicus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
Aquifex aeolicus
-
ADP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Aquifex aeolicus
O67572
gene lpxK
-
Purification (Commentary)
Commentary
Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain C41(DE3) membranes
Aquifex aeolicus
Reaction
Reaction
Commentary
Organism
ATP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate) = ADP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
ping-pong type mechanism with bi-substrate kinetics
Aquifex aeolicus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
-
737675
Aquifex aeolicus
ADP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
-
-
-
?
additional information
the interaction of residue D99 with H261 acts to increase the pKa of the imidazole moiety, which in turn serves as the catalytic base to deprotonate the 4'-hydroxyl of the DSMP substrate. The LpxK enzyme activity in vitro requires the presence of a detergent micelle and formation of a ternary LpxK-ATP/Mg2+-DSMP complex
737675
Aquifex aeolicus
?
-
-
-
-
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Aquifex aeolicus
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.00035
-
ATP
pH 8.5, 30°C, recombinant mutant D99N
Aquifex aeolicus
0.00048
-
ATP
pH 8.5, 30°C, recombinant mutant D139A; pH 8.5, 30°C, recombinant mutant E100Q
Aquifex aeolicus
0.00083
-
ATP
pH 8.5, 30°C, recombinant mutant D138A
Aquifex aeolicus
0.0013
-
ATP
pH 8.5, 30°C, recombinant mutant K51A; pH 8.5, 30°C, recombinant mutant T52A
Aquifex aeolicus
0.0015
-
ATP
pH 8.5, 30°C, recombinant mutant D99A
Aquifex aeolicus
0.0027
-
ATP
pH 8.5, 30°C, recombinant mutant E100D
Aquifex aeolicus
0.0029
-
ATP
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.0046
-
ATP
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0076
-
ATP
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.022
-
ATP
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.05
-
ATP
pH 8.5, 30°C, recombinant mutant D99E
Aquifex aeolicus
0.057
-
ATP
pH 8.5, 30°C, recombinant mutant D138N
Aquifex aeolicus
0.073
-
ATP
pH 8.5, 30°C, recombinant mutant D260A
Aquifex aeolicus
0.47
-
ATP
pH 8.5, 30°C, recombinant mutant S53A
Aquifex aeolicus
3.9
-
ATP
pH 8.5, 30°C, recombinant mutant S49A; pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
pH-dependence of active site point mutants, overview
Aquifex aeolicus
7.5
8
recombinant mutant D99A
Aquifex aeolicus
8
-
recombinant mutant H261A
Aquifex aeolicus
Cofactor
Cofactor
Commentary
Organism
Structure
ATP
the LpxK enzyme activity in vitro requires the presence of a detergent micelle and formation of a ternary LpxK-ATP/Mg2+-DSMP complex
Aquifex aeolicus
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C41(DE3) membranes
Aquifex aeolicus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
ATP
the LpxK enzyme activity in vitro requires the presence of a detergent micelle and formation of a ternary LpxK-ATP/Mg2+-DSMP complex
Aquifex aeolicus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified LpxK in complex with the ATP analogue AMPPCP in the closed catalytically competent conformation, sitting drop vapor diffusion method, mixing of 0.70 ml well solution with 0.01 ml sample solution, containing four parts of a reservoir solution consisting of 50% v/v MPD and 0.1 M HEPES, pH 7.5, and one part protein solution containing 13 mg/ml enzyme LpxK, 4.3 mM AMP-PCP, 1 mM EDTA, 0.5% w/v DDM, 540 mM NaCl, 14% v/v glycerol, and 35 mM HEPES, pH 8.0, 20°C, 1 month, or by microseeding, X-ray diffraction structure determination and analysis at 2.1 A resolution, molecular replacement using ADP-Mg2+ LpxK structure, PDB ID 4EHY, as the search model with all ligands removed, and AMP-PCP is subsequently added to the model. Purified LpxK in complex with ATP in a pre-catalytic binding state, mixing of seventeen parts of a reservoir solution consisting of 60% v/v MPD and 0.1 M HEPES, pH 7.5, and three parts protein solution containing 7.4 mg/ml enzyme LpxK, 10 mM ATP, 1 mM EDTA, 0.35% w/v DDM, 700 mM NaCl, 18.5% v/v glycerol, and 45 mM HEPES, pH 8.0, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using LpxK structure, PDB ID 4EHX, as the search model, ATP is subsequently added to the model. Purified LpxK in complex with a chloride anion in an inhibitory conformation of the nucleotide-binding P-loop, mixing of three parts of a reservoir solution consisting of 40% v/v MPD and 0.1 M HEPES, pH 7.5, and one part protein solution containing 8.3 mg/ml LpxK, 4 mM methyl 2-acetamido-2-deoxy-beta-D-glucopyranoside, 0.35% w/v DDM, 625 mM NaCl, 17% v/v glycerol, and 45 mM HEPES, pH 8.0, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement using the apo enzyme LpxK structure, PDB ID 4EHX, as search model and spherical active site density refines well as a chloride ion
Aquifex aeolicus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D138A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D138N
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D139A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D139N
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D260A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D99A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D99E
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
D99N
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E100A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E100D
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E100Q
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
H261A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
K51A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
S49A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
S53A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
T52A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
Y74A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
Ca2+
-
Aquifex aeolicus
Cu2+
-
Aquifex aeolicus
Fe3+
-
Aquifex aeolicus
Ni2+
-
Aquifex aeolicus
Zn2+
-
Aquifex aeolicus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
bi-substrate kinetics, steady-state kinetics
Aquifex aeolicus
0.0011
-
ATP
pH 8.5, 30°C, recombinant mutant D99N
Aquifex aeolicus
0.0012
-
ATP
pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
0.0013
-
ATP
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0014
-
ATP
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.0015
-
ATP
pH 8.5, 30°C, recombinant mutant K51A
Aquifex aeolicus
0.0016
-
ATP
pH 8.5, 30°C, recombinant mutant D260A
Aquifex aeolicus
0.0017
-
ATP
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.0019
-
ATP
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.002
-
ATP
pH 8.5, 30°C, recombinant mutant E100D
Aquifex aeolicus
0.0022
-
ATP
pH 8.5, 30°C, recombinant mutant T52A
Aquifex aeolicus
0.0023
-
ATP
pH 8.5, 30°C, recombinant mutant D99E
Aquifex aeolicus
0.0027
-
ATP
pH 8.5, 30°C, recombinant mutant S49A
Aquifex aeolicus
0.0028
-
ATP
pH 8.5, 30°C, recombinant mutant D99A
Aquifex aeolicus
0.0029
-
ATP
pH 8.5, 30°C, recombinant mutant E100Q
Aquifex aeolicus
0.0032
-
ATP
pH 8.5, 30°C, recombinant mutant D138N
Aquifex aeolicus
0.004
-
ATP
pH 8.5, 30°C, recombinant mutant D138A
Aquifex aeolicus
0.005
-
ATP
pH 8.5, 30°C, recombinant mutant D139A
Aquifex aeolicus
0.006
-
ATP
pH 8.5, 30°C, recombinant mutant S53A
Aquifex aeolicus
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
inner membrane
cytosol-facing, membrane binding through a hydrophobic lower face assisted by surrounding basic residues of the N-terminal core domain
Aquifex aeolicus
-
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Cl-
structural basis for anion inhibition of LpxK, modeling, overview
Aquifex aeolicus
Co2+
activates
Aquifex aeolicus
Mg2+
required, activates, optimal activity is observed at an equimolar ratio of ATP to Mg2+, with a decrease in activity at higher amounts of the divalent cation
Aquifex aeolicus
Mn2+
activates
Aquifex aeolicus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
Aquifex aeolicus
-
ADP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain C41(DE3) membranes
Aquifex aeolicus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
-
737675
Aquifex aeolicus
ADP + (2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucosaminyl)-(1->6)-(2-N,3-O-bis[(3R)-3-hydroxytetradecanoyl]-alpha-D-glucosaminyl phosphate)
-
-
-
?
additional information
the interaction of residue D99 with H261 acts to increase the pKa of the imidazole moiety, which in turn serves as the catalytic base to deprotonate the 4'-hydroxyl of the DSMP substrate. The LpxK enzyme activity in vitro requires the presence of a detergent micelle and formation of a ternary LpxK-ATP/Mg2+-DSMP complex
737675
Aquifex aeolicus
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Aquifex aeolicus
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.00035
-
ATP
pH 8.5, 30°C, recombinant mutant D99N
Aquifex aeolicus
0.00048
-
ATP
pH 8.5, 30°C, recombinant mutant D139A; pH 8.5, 30°C, recombinant mutant E100Q
Aquifex aeolicus
0.00083
-
ATP
pH 8.5, 30°C, recombinant mutant D138A
Aquifex aeolicus
0.0013
-
ATP
pH 8.5, 30°C, recombinant mutant K51A; pH 8.5, 30°C, recombinant mutant T52A
Aquifex aeolicus
0.0015
-
ATP
pH 8.5, 30°C, recombinant mutant D99A
Aquifex aeolicus
0.0027
-
ATP
pH 8.5, 30°C, recombinant mutant E100D
Aquifex aeolicus
0.0029
-
ATP
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.0046
-
ATP
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0076
-
ATP
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.022
-
ATP
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.05
-
ATP
pH 8.5, 30°C, recombinant mutant D99E
Aquifex aeolicus
0.057
-
ATP
pH 8.5, 30°C, recombinant mutant D138N
Aquifex aeolicus
0.073
-
ATP
pH 8.5, 30°C, recombinant mutant D260A
Aquifex aeolicus
0.47
-
ATP
pH 8.5, 30°C, recombinant mutant S53A
Aquifex aeolicus
3.9
-
ATP
pH 8.5, 30°C, recombinant mutant S49A; pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
pH-dependence of active site point mutants, overview
Aquifex aeolicus
7.5
8
recombinant mutant D99A
Aquifex aeolicus
8
-
recombinant mutant H261A
Aquifex aeolicus
General Information
General Information
Commentary
Organism
evolution
kinase LpxK is a member of the P-loop containing nucleoside triphosphate hydrolase superfamily. The active site Walker A (P-loop) and Walker B (Mg2+-binding) motifs are common to all P-loop kinase family members
Aquifex aeolicus
metabolism
the sixth step in the lipid A biosynthetic pathway involves phosphorylation of the tetraacyldisaccharide-1-phosphate (DSMP) intermediate by the cytosol-facing inner membrane kinase LpxK
Aquifex aeolicus
additional information
apparent steady-state kinetic parameters for LpxK activity support the formation of a ternary LpxK-ATP/Mg2+-DSMP complex. In its closed catalytically competent form, the C-terminal domain of LpxK undergoes a hinge motion to close around the nucleotide substrate upon binding. Active sites of ATP-bound enzyme LpxK are in the open form, modeling, overview
Aquifex aeolicus
physiological function
in the reaction catalyzed by LpxK in Kdo2-lipid A biosynthesis, enzyme LpxK is responsible for the phosphorylation of the 4'-hydroxyl of tetraacyldisaccharide-1-phosphate (DSMP)
Aquifex aeolicus
General Information (protein specific)
General Information
Commentary
Organism
evolution
kinase LpxK is a member of the P-loop containing nucleoside triphosphate hydrolase superfamily. The active site Walker A (P-loop) and Walker B (Mg2+-binding) motifs are common to all P-loop kinase family members
Aquifex aeolicus
metabolism
the sixth step in the lipid A biosynthetic pathway involves phosphorylation of the tetraacyldisaccharide-1-phosphate (DSMP) intermediate by the cytosol-facing inner membrane kinase LpxK
Aquifex aeolicus
additional information
apparent steady-state kinetic parameters for LpxK activity support the formation of a ternary LpxK-ATP/Mg2+-DSMP complex. In its closed catalytically competent form, the C-terminal domain of LpxK undergoes a hinge motion to close around the nucleotide substrate upon binding. Active sites of ATP-bound enzyme LpxK are in the open form, modeling, overview
Aquifex aeolicus
physiological function
in the reaction catalyzed by LpxK in Kdo2-lipid A biosynthesis, enzyme LpxK is responsible for the phosphorylation of the 4'-hydroxyl of tetraacyldisaccharide-1-phosphate (DSMP)
Aquifex aeolicus
Other publictions for EC 2.7.1.130
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)
738621
Emptage
Structural basis of lipid bind ...
Aquifex aeolicus
J. Biol. Chem.
289
24059-24068
2014
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737675
Emptage
Mechanistic characterization o ...
Aquifex aeolicus
Biochemistry
52
2280-2290
2013
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723650
Emptage
Crystal structure of LpxK, the ...
Aquifex aeolicus
Proc. Natl. Acad. Sci. USA
109
12956-12961
2012
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640807
Garrett
Identification of the gene enc ...
Escherichia coli
J. Biol. Chem.
272
21855-21864
1997
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640806
Hampton
Lipid A 4-kinase from Escheric ...
Escherichia coli
Methods Enzymol.
209
466-475
1992
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Ray
The biosynthesis of gram-negat ...
Escherichia coli
J. Biol. Chem.
262
1122-1128
1987
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