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show all sequences of 2.7.1.130

Structural basis of lipid binding for the membrane-embedded tetraacyldisaccharide-1-phosphate 4-kinase LpxK

Emptage, R.P.; Tonthat, N.K.; York, J.D.; Schumacher, M.A.; Zhou, P.; J. Biol. Chem. 289, 24059-24068 (2014)

Data extracted from this reference:

Application
Application
Commentary
Organism
drug development
the enzyme structure provides a structural template for designing antibiotics and knowledge of catalysis at the membrane interface
Aquifex aeolicus
Cloned(Commentary)
Commentary
Organism
gene lpxK, complementation of an Escherichia coli lpxK chromosomal knockout mutant strain W3110 with expression of Aquifex aeolicus gene lpxK, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C41(DE3) membranes
Aquifex aeolicus
Crystallization (Commentary)
Crystallization
Organism
purified enzyme LpxK in complex with lipid IVA, X-ray diffraction structure determination and analysis at 3.5 A
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
E100A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E172A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
H143A
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
additional information
steady-state kinetic analysis of multiple point mutants of the lipid-binding pocket pinpoints critical residues involved in substrate binding, and construction of two N-terminal helix truncated forms of LpxK, one in which amino acids 2-12 are removed, DELTA12LpxK, and another in which amino acids 2-29 are removed, DELTA29LpxK
Aquifex aeolicus
N43A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
Q142A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
R119A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
R171A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
R72A
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
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.007
-
(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)
pH 8.5, 30°C, recombinant mutant D138A; pH 8.5, 30°C, recombinant mutant D138N; pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
0.009
-
(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)
pH 8.5, 30°C, recombinant mutant N43A
Aquifex aeolicus
0.0097
-
(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)
pH 8.5, 30°C, recombinant mutant R119A
Aquifex aeolicus
0.014
-
(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)
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.015
-
(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)
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0174
-
(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)
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.018
-
(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)
pH 8.5, 30°C, recombinant mutant E172A
Aquifex aeolicus
0.028
-
(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)
pH 8.5, 30°C, recombinant mutant Q142A; pH 8.5, 30°C, recombinant mutant R72A
Aquifex aeolicus
0.046
-
(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)
pH 8.5, 30°C, recombinant mutant D139A
Aquifex aeolicus
0.048
-
(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)
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.05
-
(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)
pH 8.5, 30°C, recombinant mutant H143A
Aquifex aeolicus
0.128
-
(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)
pH 8.5, 30°C, recombinant mutant R171A
Aquifex aeolicus
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
the enzyme is an integral membrane protein
Aquifex aeolicus
16020
-
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 by solubilization from membranes with Triton X-100, and ultracentrifugation
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)
-
738621
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
lipid IVA is bound in the putative lipid-binding pocket on the underside of the N-terminal enzyme domain. The L4 loop, which includes the Walker B motif, appears to contain important residues for binding the lipid substrate, lipid IVA binding structures of wild-type and mutant enzymes, overview
738621
Aquifex aeolicus
?
-
-
-
-
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.00051
-
(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)
pH 8.5, 30°C, recombinant mutant D139A
Aquifex aeolicus
0.0011
-
(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)
pH 8.5, 30°C, recombinant mutant D138A
Aquifex aeolicus
0.00123
-
(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)
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.0031
-
(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)
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0063
-
(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)
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.0138
-
(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)
pH 8.5, 30°C, recombinant mutant R171A
Aquifex aeolicus
0.028
-
(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)
pH 8.5, 30°C, recombinant mutant D138N
Aquifex aeolicus
0.031
-
(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)
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.036
-
(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)
pH 8.5, 30°C, recombinant mutant R119A
Aquifex aeolicus
0.16
-
(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)
pH 8.5, 30°C, recombinant mutant H143A
Aquifex aeolicus
0.35
-
(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)
pH 8.5, 30°C, recombinant mutant Q142A
Aquifex aeolicus
0.36
-
(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)
pH 8.5, 30°C, recombinant mutant N43A
Aquifex aeolicus
0.39
-
(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)
pH 8.5, 30°C, recombinant mutant R72A
Aquifex aeolicus
0.7
-
(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)
pH 8.5, 30°C, recombinant mutant E172A
Aquifex aeolicus
2.4
-
(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)
pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
Application (protein specific)
Application
Commentary
Organism
drug development
the enzyme structure provides a structural template for designing antibiotics and knowledge of catalysis at the membrane interface
Aquifex aeolicus
Cloned(Commentary) (protein specific)
Commentary
Organism
gene lpxK, complementation of an Escherichia coli lpxK chromosomal knockout mutant strain W3110 with expression of Aquifex aeolicus gene lpxK, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain C41(DE3) membranes
Aquifex aeolicus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified enzyme LpxK in complex with lipid IVA, X-ray diffraction structure determination and analysis at 3.5 A
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
E100A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
E172A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
H143A
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
additional information
steady-state kinetic analysis of multiple point mutants of the lipid-binding pocket pinpoints critical residues involved in substrate binding, and construction of two N-terminal helix truncated forms of LpxK, one in which amino acids 2-12 are removed, DELTA12LpxK, and another in which amino acids 2-29 are removed, DELTA29LpxK
Aquifex aeolicus
N43A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
Q142A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
R119A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
R171A
site-directed mutagenesis, the mutant shows altered activity compared to the wild-type enzyme
Aquifex aeolicus
R72A
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
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.007
-
(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)
pH 8.5, 30°C, recombinant mutant D138A; pH 8.5, 30°C, recombinant mutant D138N; pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
0.009
-
(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)
pH 8.5, 30°C, recombinant mutant N43A
Aquifex aeolicus
0.0097
-
(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)
pH 8.5, 30°C, recombinant mutant R119A
Aquifex aeolicus
0.014
-
(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)
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.015
-
(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)
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0174
-
(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)
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.018
-
(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)
pH 8.5, 30°C, recombinant mutant E172A
Aquifex aeolicus
0.028
-
(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)
pH 8.5, 30°C, recombinant mutant Q142A; pH 8.5, 30°C, recombinant mutant R72A
Aquifex aeolicus
0.046
-
(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)
pH 8.5, 30°C, recombinant mutant D139A
Aquifex aeolicus
0.048
-
(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)
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.05
-
(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)
pH 8.5, 30°C, recombinant mutant H143A
Aquifex aeolicus
0.128
-
(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)
pH 8.5, 30°C, recombinant mutant R171A
Aquifex aeolicus
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
membrane
the enzyme is an integral membrane protein
Aquifex aeolicus
16020
-
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 by solubilization from membranes with Triton X-100, and ultracentrifugation
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)
-
738621
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
lipid IVA is bound in the putative lipid-binding pocket on the underside of the N-terminal enzyme domain. The L4 loop, which includes the Walker B motif, appears to contain important residues for binding the lipid substrate, lipid IVA binding structures of wild-type and mutant enzymes, overview
738621
Aquifex aeolicus
?
-
-
-
-
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.00051
-
(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)
pH 8.5, 30°C, recombinant mutant D139A
Aquifex aeolicus
0.0011
-
(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)
pH 8.5, 30°C, recombinant mutant D138A
Aquifex aeolicus
0.00123
-
(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)
pH 8.5, 30°C, recombinant mutant E100A
Aquifex aeolicus
0.0031
-
(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)
pH 8.5, 30°C, recombinant mutant H261A
Aquifex aeolicus
0.0063
-
(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)
pH 8.5, 30°C, recombinant mutant D139N
Aquifex aeolicus
0.0138
-
(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)
pH 8.5, 30°C, recombinant mutant R171A
Aquifex aeolicus
0.028
-
(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)
pH 8.5, 30°C, recombinant mutant D138N
Aquifex aeolicus
0.031
-
(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)
pH 8.5, 30°C, recombinant mutant Y74A
Aquifex aeolicus
0.036
-
(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)
pH 8.5, 30°C, recombinant mutant R119A
Aquifex aeolicus
0.16
-
(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)
pH 8.5, 30°C, recombinant mutant H143A
Aquifex aeolicus
0.35
-
(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)
pH 8.5, 30°C, recombinant mutant Q142A
Aquifex aeolicus
0.36
-
(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)
pH 8.5, 30°C, recombinant mutant N43A
Aquifex aeolicus
0.39
-
(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)
pH 8.5, 30°C, recombinant mutant R72A
Aquifex aeolicus
0.7
-
(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)
pH 8.5, 30°C, recombinant mutant E172A
Aquifex aeolicus
2.4
-
(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)
pH 8.5, 30°C, recombinant wild-type enzyme
Aquifex aeolicus
General Information
General Information
Commentary
Organism
additional information
structural and kinetic studies reveal the molecular basis of lipid binding, overview. The LpxK active site recognizes the lipid's glucosamine/phosphate headgroups and only accommodates disaccharides. Steady-state kinetic analysis of multiple point mutants of the lipid-binding pocket pinpoints critical residues involved in substrate binding, and characterization of N-terminal helix truncation mutants uncovers the role of this substructure as a hydrophobic membrane anchor
Aquifex aeolicus
physiological function
enzyme LpxK is an essential membrane-bound kinase in the lipid A biosynthetic pathway. In Gram-negative bacteria, lipidA is the hydrophobic anchor of lipopolysaccharide, which makes up the outer leaflet of the asymmetric outer membrane of these organisms. This acylated disaccharide of glucosamine plays an important role in eliciting an immunogenic response to bacterial pathogens and is essential to the survival of the vast majority of these microbes
Aquifex aeolicus
General Information (protein specific)
General Information
Commentary
Organism
additional information
structural and kinetic studies reveal the molecular basis of lipid binding, overview. The LpxK active site recognizes the lipid's glucosamine/phosphate headgroups and only accommodates disaccharides. Steady-state kinetic analysis of multiple point mutants of the lipid-binding pocket pinpoints critical residues involved in substrate binding, and characterization of N-terminal helix truncation mutants uncovers the role of this substructure as a hydrophobic membrane anchor
Aquifex aeolicus
physiological function
enzyme LpxK is an essential membrane-bound kinase in the lipid A biosynthetic pathway. In Gram-negative bacteria, lipidA is the hydrophobic anchor of lipopolysaccharide, which makes up the outer leaflet of the asymmetric outer membrane of these organisms. This acylated disaccharide of glucosamine plays an important role in eliciting an immunogenic response to bacterial pathogens and is essential to the survival of the vast majority of these microbes
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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15
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737675
Emptage
Mechanistic characterization o ...
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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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8
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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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640805
Ray
The biosynthesis of gram-negat ...
Escherichia coli
J. Biol. Chem.
262
1122-1128
1987
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