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Information on EC 2.3.1.129 - acyl-[acyl-carrier-protein]-UDP-N-acetylglucosamine O-acyltransferase and Organism(s) Escherichia coli and UniProt Accession P0A722
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EC Tree
2.3.1.129 acyl-[acyl-carrier-protein]-UDP-N-acetylglucosamine O-acyltransferaseIUBMB Comments
Involved with EC 2.4.1.182, lipid-A-disaccharide synthase, and EC 2.7.1.130, tetraacyldisaccharide 4'-kinase, in the biosynthesis of the phosphorylated glycolipid, Lipid A, in the outer membrane of Gram-negative bacteria.
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Word Map
- 2.3.1.129
- udp-glcnac
- lipopolysaccharide
-
left-handed
- acyl-acp
- endotoxin
- drug development
-
beta-helix
-
beta-helical
-
homotrimer
-
udp-3-o-r-3-hydroxymyristoyl-glcnac
-
raetz
- medicine
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
Reaction Schemes
Synonyms
udp-n-acetylglucosamine acyltransferase, lilpxa, udp-n-acetylglucosamine 3-o-acyltransferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
acyltransferase, uridine diphosphoacetylglucosamine
-
-
-
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type II acyl carrier protein-dependent UDP-N-acetylglucosamine acyltransferase
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-
UDP-N-acetylglucosamine acyltransferase
uridine diphosphoacetylglucosamine acyltransferase
-
-
-
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UDP-N-acetylglucosamine acyltransferase
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-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a (3R)-3-hydroxyacyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine = an [acyl-carrier protein] + a UDP-3-O-[(3R)-3-hydroxyacyl]-N-acetyl-alpha-D-glucosamine
a (3R)-3-hydroxyacyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine = an [acyl-carrier protein] + a UDP-3-O-[(3R)-3-hydroxyacyl]-N-acetyl-alpha-D-glucosamine
reaction mechanism, acylation occurs via nucleophilic attack of deprotonated UDP-GlcNAc on the acyl donor in a general base-catalyzed mechanism involving a catalytic dyad of His125 and Asp126, His125, the general base, interacts with the 3'-hydroxyl group of UDP-GlcNAc to generate the nucleophile, the Asp126 side-chain accepts a hydrogen bond from His125 and helps orient the general base to participate in catalysis
a (3R)-3-hydroxyacyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine = an [acyl-carrier protein] + a UDP-3-O-[(3R)-3-hydroxyacyl]-N-acetyl-alpha-D-glucosamine
reaction mechanism, acylation occurs via nucleophilic attack of deprotonated UDP-GlcNAc on the acyl donor in a general base-catalyzed mechanism involving a catalytic dyad of His125 and Asp126, His125, the general base, interacts with the 3'-hydroxyl group of UDP-GlcNAc to generate the nucleophile, the Asp126 side-chain accepts a hydrogen bond from His125 and helps orientate the general base to participate in catalysis
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Acyl group transfer
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PATHWAY SOURCE
PATHWAYS
BRENDA
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-, -, -, -, -, -
SYSTEMATIC NAME
IUBMB Comments
(3R)-3-hydroxyacyl-[acyl-carrier protein]:UDP-N-acetyl-alpha-D-glucosamine 3-O-(3-hydroxyacyl)transferase
Involved with EC 2.4.1.182, lipid-A-disaccharide synthase, and EC 2.7.1.130, tetraacyldisaccharide 4'-kinase, in the biosynthesis of the phosphorylated glycolipid, Lipid A, in the outer membrane of Gram-negative bacteria.
CAS REGISTRY NUMBER
COMMENTARY
105843-69-4
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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
(R)-3-hydroxydodecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxydodecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
?
(R)-3-hydroxymyristoyl-[acyl carrier protein] + UDP-N-acetylglucosamine
[acyl-carrier protein] + UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine
first step in lipid A biosynthesis
-
-
?
(R)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxymyristoyl)-N-acetyl-alpha-D-glucosamine
first step in lipid A biosynthesis
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
R-3-hydroxydecanoyl-[acyl carrier protein] + UDP-N-acetylglucosamine
[acyl-carrier protein] + UDP-3-O-hydroxydecanoyl-N-acetylglucosamine
first step in lipid A biosynthesis
-
-
?
(R)-3-hydroxydodecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxydodecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R)-3-hydroxyhexadecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxyhexadecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucopyranose
[acyl-carrier protein] + UDP-3-N-(3-hydroxytetradecanoyl)-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucopyranose
-
only in vitro
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-butyrylglucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-butyrylglucosamine
-
transacylation at 8% the rate of the reaction with UDP-N-acetylglucosamine
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-propionyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-propionyl-alpha-D-glucosamine
-
transacylation at 22% the rate of the reaction with UDP-N-acetylglucosamine
-
-
?
(R,S)-3-hydroxydodecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxydodecanoyl)-N-acetyl-alpha-D-glucosamine
(R,S)-3-hydroxylauroyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxylauroyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R,S)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxymyristoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R,S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
(S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
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transacylation at 7% the rate of (R)-enantiomer
-
-
?
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
LpxA is essential for the growth of Escherichia coli, and is important in lipid A biosynthesis, overview
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-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
the enzyme is responsible for the first step of lipid A biosynthesis, lipid A is an integral component of the lipopolysaccharide that forms the selective and protective outer monolayer of Gram-negative bacteria, and is essential for bacterial growth and viability
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
i.e. (R)-3-hydroxymyristoyl-[acyl-carrier protein], the enzyme catalyzes the first step of lipid A biosynthesis, the transfer of the (R)-3-hydroxyacyl chain from (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] to the glucosamine 3-OH group of UDP-GlcNAc
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
substrate binding site structure with prominent hydrophilic interactions between highly conserved clusters of residues Asn198, Glu200, Arg204 and Arg205 with UDP, and Asp74, His125, His144 and Gln161 with the GlcNAc moiety, these interactions serve to bind and orient the substrate for catalysis, overview
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-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
substrate binding site structure with prominent hydrophilic interactions between highly conserved clusters of residues Asn198, Glu200, Arg204 and Arg205 with UDP, and Asp74, His125, His144 and Gln161 with the GlcNAc moiety, these interactions serve to bind and orientate the substrate for catalysis, overview
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-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
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essential for cell growth of gram-negative bacteria
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
involved with EC 2.4.1.182 and 2.7.1.130 in the biosynthesis of the phosphorylated glycolipid and outer membrane component, Lipid A
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
initial reaction in lipid A biosynthesis
-
-
?
(R,S)-3-hydroxydodecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxydodecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R,S)-3-hydroxydodecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxydodecanoyl)-N-acetyl-alpha-D-glucosamine
-
i.e. 3-hydroxylauroyl-[acyl-carrier-protein], poor substrate
-
-
?
(R,S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R,S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
r
(R,S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
no substrates are UDPglucosamine, UDP-N-(R)-3-hydroxymyristoylglucosamine, ADP-N-acetylglucosamine, GDP-N-acetylglucosamine, CDP-N-acetylglucosamine, (R)-3-hydroxytetradecanoyl-CoA, or palmitoyl-CoA, myristoyl-[acyl-carrier-protein], and palmitoyl-[acyl-carrier-protein]
-
-
?
(R,S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
no substrates are UDPglucosamine, UDP-N-(R)-3-hydroxymyristoylglucosamine, ADP-N-acetylglucosamine, GDP-N-acetylglucosamine, CDP-N-acetylglucosamine, (R)-3-hydroxytetradecanoyl-CoA, or palmitoyl-CoA, myristoyl-[acyl-carrier-protein], and palmitoyl-[acyl-carrier-protein]
-
-
r
(R,S)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
high specificity with respect to acyl-donor, equilibrium constant favors thioester acyl carrier substrate
-
-
r
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
r
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
assay is optimal at 10 nM LpxA
-
-
?
additional information
?
-
nucleotide substrate recognition by LpxA, overview
-
-
?
additional information
?
-
-
nucleotide substrate recognition by LpxA, overview
-
-
?
additional information
?
-
-
substrate specificity, no activity with UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucopyranose due to lack of genes gnnA and gnnB, lipid A structure overview
-
-
?
additional information
?
-
-
development of a fluorescence polarization assay, detecting displacement of a fluorescently labeled peptide inhibitor based on the previously reported inhibitor peptide 920 by active site ligands, for high-throughput screening of LpxA competitve inhibitors, overview
-
-
?
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
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
LpxA is essential for the growth of Escherichia coli, and is important in lipid A biosynthesis, overview
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
the enzyme is responsible for the first step of lipid A biosynthesis, lipid A is an integral component of the lipopolysaccharide that forms the selective and protective outer monolayer of Gram-negative bacteria, and is essential for bacterial growth and viability
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
essential for cell growth of gram-negative bacteria
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
involved with EC 2.4.1.182 and 2.7.1.130 in the biosynthesis of the phosphorylated glycolipid and outer membrane component, Lipid A
-
-
?
(R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
[acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
initial reaction in lipid A biosynthesis
-
-
?
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
?
an (R)-3-hydroxytetradecanoyl-[acyl-carrier protein] + UDP-N-acetyl-alpha-D-glucosamine
an [acyl-carrier protein] + UDP-3-O-(3-hydroxytetradecanoyl)-N-acetyl-alpha-D-glucosamine
-
-
-
-
r
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FITC-P920
-
i.e. FITC-(beta)SSGWMLDPIAGKWSRNH2, fluoresecent-labeled peptide
peptide 920
-
i.e. pentadecapeptide 920, with C-terminus WMLDPIAGKWSR, competitive versus peptide inhibitors derived from its own C-terminal sequence
-
RJPXD33
-
i.e. TNLYMLPKWDIP-NH2, a peptide identified from a phage-bound random peptide library screen uing Escherichia oli strain XL-1 Blue, binds to UDP-3-O-(R-3-hydroxyacyl)GlcN N-acyltransferase, LpxD, and UDP-N-acetylglucosamine acyltransferase, LpxA. RJPXD33 binds to LpxA in a competitive fashion with P920
TAMRA peptide
-
ability of the TAMRA-peptide to inhibit the catalytic activity
-
UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine
-
competitive versus TAMRA peptide
peptide 920
pentadecapeptide NH2-SSGWMLDPIAGKWSR-COOH, the enzyme binds peptide 920 with three peptides, each of which adopts a beta-hairpin conformation, bound per LpxA trimer, the peptides are located at the interfaces of adjacent subunits in the vicinity of the three active sites, each peptide interacts with residues from both adjacent subunits, noncovalent interaction and binding structure, overview
-
additional information
-
no inhibition by N-ethylmaleimide, UDP-2,3-diacyl-N-glucosamine, lipid X, lipid IVA, 3-deoxy-D-mannooctulosonate, octyl-beta-D-glucoside
-
additional information
-
no inhibition by N-ethylmaleimide, UDP-2,3-diacyl-N-glucosamine, lipid X, lipid IVA, 3-deoxy-D-mannooctulosonate, octyl-beta-D-glucoside
-
additional information
-
no inhibition by N-ethylmaleimide, UDP-2,3-diacyl-N-glucosamine, lipid X, lipid IVA, 3-deoxy-D-mannooctulosonate, octyl-beta-D-glucoside
-
additional information
-
development of a fluorescence polarization assay, detecting displacement of a fluorescently labeled peptide inhibitor, based on inhibitor peptide 920 C-terminal 12 residues WMLDPIAGKWSR, by active site ligands, for high-throughput screening of LpxA competitve inhibitors, overview. Abilities of several ligands to compete with TAMRA peptide for binding to Escherichia coli LpxA. Acyl carrier protein does not significantly affect the inhibitory potencies
-
additional information
-
measurement of binding of fluorescein-labeled peptide to acyltransferase by measuring the changes in fluorescence polarization, KD values, overview. No inhibition of LpxA by SENNFMLPLLPL-NH2, i.e. peptide RJPXD34
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
Michaelis-Menten steady-state kinetics of LpxA, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00005
peptide 920
peptide 920 disrupts LpxA interactions with the R-3-hydroxymyristoyl-ACP substrate
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00014
HM-UNAG peptide
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in absence of holo-ACP
-
0.000238
HM-UNAG peptide
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in presence of holo-ACP
-
0.047
HMA peptide
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in absence of holo-ACP
-
0.063
HMA peptide
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in presence of holo-ACP
-
0.00006
peptide 920
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide
-
0.00051
peptide 920
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in presence of holo-ACP
-
0.0006
peptide 920
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus 0.001 mM UNAG peptide
-
0.00073
peptide 920
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in absence of holo-ACP
-
0.000923
peptide 920
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus 5 mM UNAG peptide
-
5.8
UNAG peptide
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in absence of holo-ACP
-
6.6
UNAG peptide
Escherichia coli
-
forward reaction, pH 7.0, 22°C, versus TAMRA peptide, in presence of holo-ACP
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.000006
-
S)-3-hydroxydecanoyl-[acyl-carrier-protein] as substrate, pTO10 and pTO105
0.000007
-
S)-3-hydroxydodecanoyl-[acyl-carrier-protein] as substrate, pTO10
0.00006
-
S)-3-hydroxytetradecanoyl-[acyl-carrier-protein] as substrate, pTO101
0.000068
-
S)-3-hydroxytetradecanoyl-[acyl-carrier-protein] as substrate, pTO104
0.00007
0.000087
-
S)-3-hydroxytetradecanoyl-[acyl-carrier-protein] as substrate, pTO103
0.0002
-
S)-3-hydroxytetradecanoyl-[acyl-carrier-protein] as substrate, pTO102
0.000347
-
S)-3-hydroxytetradecanoyl-[acyl-carrier-protein] as substrate, pTO10
0.00053
-
S)-3-hydroxytetradecanoyl-[acyl-carrier-protein] as substrate, pTO102
additional information
-
substrate specificity with different acyl acceptors, activity of the recombinant enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
the enzymes in early lipid A biosynthesis are highly conserved in gram-negative pathogens. LpxA and LpxD are structurally homologous possessing the unique left-handed beta-helix fold, which stems from an extensive hexapeptide repeat motif in their respective primary amino acid sequences
metabolism
physiological function
additional information
-
UDP-N-acetylglucosamine 3-O-acyltransferase is a protein with a left-handed parallel beta-helix, which is a natural nanotube associated with unusual high stability, molecular dynamics simulations using LpxA crystal structure, PDB ID 1LXA, and formation of a dynamical cross-correlation map, overview. Construction of the unfolding conformational energy landscape identifies the probable intermediates that can appear in the unfolding pathway of the protein, unfolding kinetics of the three-stranded beta-sheet protein, overview
metabolism
-
acyltransferases LpxA and LpxD catalyze functionally similar acylations of their respective UDP-glucosamine-based substrates using R-3-hydroxymyristoyl-ACP as the acyl donor in the Kdo2-lipid A biosynthesis in Escherichia coli
metabolism
-
LpxA is the first enzyme in the biosynthetic pathway for the Lipid A component of the outer membrane lipopolysaccharide
metabolism
-
UDP-N-acetylglucosamine acyltransferase, LpxA, catalyzes the first step of lipid A biosynthesis
physiological function
-
acyl-carrier protein (ACP), a small 8-10 kDa component of the type II dissociated fatty acid synthase system, of Vibrio harveyi interacts with LpxA and induces ACP-folding
physiological function
-
LpxA catalyzes the condensation of UDP N-acetylglucosamine with R-3-hydroxymyristic acid from R-3-hydroxymyristoyl-acyl carrier protein in the biosynthesis of lipopolysaccharide
physiological function
-
UDP-GlcNAc is acylated at the 3-hydroxyl through a thermodynamically unfavorable reaction catalyzed by the type II acyl carrier protein-dependent UDP-N-acetylglucosamine acyltransferase, LpxA, first step in biosynthesis of lipid A and essential for survival among Gram-negative bacteria that synthesize lipopolysaccharide. Lipopolysaccharide contains the glycolipid lipid A, which anchors it to the membrane through fatty acyl chains, a core polysaccharide region, and an O-antigen repeat. Lipid A is an essential moiety necessary for survival of the bacterium and further plays a crucial role in natural antibacterial resistance and bacterial sepsis
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
trimer
trimer
trimer
homotrimer, LpxA contains an unusual, left-handed parallel beta-helix fold, crystal structure analysis
trimer
homotrimer, the active site lies within a positively charged cleft formed at the subunit-subunit interface
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
by the sitting-drop, vapour-diffusion method using 0.00l ml of protein solution
by using the hanging drop vapor diffusion method, in the presence of a 25-fold molar excess of either UDP-3-O-(R-3-hydroxymyristoyl)-GlcNAc or UDP-3-O-(R-3-hydroxydecanoyl)-GlcNAc, structures show how LpxA selects for 14-carbon R-3-hydroxyacyl chains and reveal two modes of UDP binding
in complex with inhibitor peptide WMLDPIAGKWSR, to 1.6 A resolution. The peptide is located at the interface of each adjacent subunit and interacts with residues from both sides. It occupies part of the ACP binding site
purified enzyme LpxA in a complex with inhibitor peptide 920, 20 mg/ml protein in solution with a 25fold molar excess of peptide 920 of 12.5 mM, crystal growth at 18°C, hanging drop vapor diffusion method, mixing of 0.002 ml protein solution with 0.002 ml of 0.81.8 M phosphate buffer, pH 6.36.9, and 30-35% DMSO, about 2 weeks, X-ray diffraction structure determination and analysis at 1.8 A resolution, molecular replacement using crystal structure PDB ID code 1LXA, determined at 2.6 A resolution
structure in complex with peptide RJPXD33 at 1.9 A resolution. Results suggest that the peptide binds in a unique modality that mimics (R)-beta-hydroxyacyl pantetheine binding to LpxA. REsidue H160 changes its conformation upon binding of peptide. Overlay of the LpxA RJPXD33 structure with LpxD, EC 2.31.191, identifies a complementary peptide binding pocket within LpxD and serves as a model for characterization of RJPXD33 binding to LpxD
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
G189S
Escherichia coli strain SM101 is deficient in LpxA activity due to a G189S inactivating mutation. Enzymatic activity is restored when the mutant strain is transformed with a wild-type bearing plasmid is inactive
G52L/R58L
residue tolerance in the beta-helical domain: mutation is tolerated
G52V/R58V
residue tolerance in the beta-helical domain: mutation is tolerated
G52W/R58W
residue tolerance in the beta-helical domain: mutation is tolerated
G57P/N51P
residue tolerance in the beta-helical domain: mutation is not tolerated
H125A
H125 is an important residue at the active site its mutation eliminates activity. Mutant shows significant growth reduction after introduction into Escherichia coli strain SM101 bearing a defective LpxA gene (G189S), and under novobiocin supplementation
H53I/D59I
residue tolerance in the beta-helical domain: mutation is tolerated
H53V/D59V
residue tolerance in the beta-helical domain: mutation is tolerated
I20R
mutation in the hydrophobic residue in the beta-helical core located in rung 2: LpxA is active
I2R
mutation in the hydrophobic residue in the beta-helical core located in rung 1: LpxA is active
I38R
mutation in the hydrophobic residue in the beta-helical core located in rung 3: LpxA is partially active
I56A
mutation in the hydrophobic core of the beta-helical domain: LpxA activity is not significantly affected compared to wild-type
I56D/I62D
residue tolerance in the beta-helical domain: mutation is not tolerated
I56E/I62E
residue tolerance in the beta-helical domain: mutation is not tolerated
I56G
mutation in the hydrophobic core of the beta-helical domain: LpxA activity is decreased compared to wild-type
I56G/I62G
residue tolerance in the beta-helical domain: mutation is tolerated
I56H/I62H
residue tolerance in the beta-helical domain: mutation is not tolerated
I56K/I62K
residue tolerance in the beta-helical domain: mutation is not tolerated
I56N
mutation in the hydrophobic core of the beta-helical domain: LpxA activity is decreased compared to wild-type
I56N/I62N
residue tolerance in the beta-helical domain: mutation is tolerated
I56P/I62P
residue tolerance in the beta-helical domain: mutation is not tolerated
I56Q
mutation in the hydrophobic core of the beta-helical domain: LpxA activity is decreased compared to wild-type
I56Q/I62Q
residue tolerance in the beta-helical domain: mutation is tolerated
I56R
I56R/I62R
residue tolerance in the beta-helical domain: mutation is not tolerated
I56S/I62S
residue tolerance in the beta-helical domain: mutation is tolerated
I56W/I62W
residue tolerance in the beta-helical domain: mutation is not tolerated
I56Y/I62Y
residue tolerance in the beta-helical domain: mutation is not tolerated
I86R
K55D/E61D
residue tolerance in the beta-helical domain: mutation is tolerated
K55P/E61P
residue tolerance in the beta-helical domain: mutation is not tolerated
P10A/P28A/P34A/P183A
proline mutation at the turn region of the beta-helical domain: mutant shows lower activity compared to wild-type
P28A/P34A
proline mutation at the turn region of the beta-helical domain: mutant shows greater activity than the P10A/P28A/P34A/P183A mutant
T54D/N60D
residue tolerance in the beta-helical domain: mutation is not tolerated
T54E/N60E
residue tolerance in the beta-helical domain: mutation is not tolerated
T54G/N60G
residue tolerance in the beta-helical domain: mutation is tolerated
T54H/N60H
residue tolerance in the beta-helical domain: mutation is not tolerated
T54K/N60K
residue tolerance in the beta-helical domain: mutation is not tolerated
T54M/N60M
residue tolerance in the beta-helical domain: mutation is tolerated
T54P/N60P
residue tolerance in the beta-helical domain: mutation is not tolerated
T54Q/N60Q
residue tolerance in the beta-helical domain: mutation is tolerated
T54R/N60R
residue tolerance in the beta-helical domain: mutation is not tolerated
T54W/N60W
residue tolerance in the beta-helical domain: mutation is not tolerated
T54Y/N60Y
residue tolerance in the beta-helical domain: mutation is not tolerated
V111R
mutation in the hydrophobic residue in the beta-helical core located in rung 6: LpxA is inactive
V129R
mutation in the hydrophobic residue in the beta-helical core located in rung 7: LpxA is inactive
Y66F/Y77F/Y219F/Y223F/Y243H
all but one tyrosine residues are mutated. Mutant shows growth similar to wild-type after introduction into Escherichia coli strain SM101 bearing a defective LpxA gene (G189S), and under novobiocin supplementation
additional information
I56R
mutation in the hydrophobic core of the beta-helical domain: LpxA activity is completely abolished compared to wild-type
I56R
mutation in the hydrophobic residue in the beta-helical core located in rung 4: LpxA is inactive
I86R
mutation in the hydrophobic residue in the beta-helical core located in rung 5: LpxA is inactive
I86R
residue is located in the fifth rung of the beta-helical domain in the hydrophobic core of a beta helix, mutant is improperly folded, destabilized and its enzymatic activity is decreased. Mutant shows significant growth reduction after introduction into Escherichia coli strain SM101 bearing a defective LpxA gene (G189S), and under novobiocin supplementation
additional information
a recombinant prion protein-LpxA protein is generated, in which a PrP fragment that is thought to be essential for the conformational conversion is incorporated into the beta-helical domain of LpxA. Partial Lpxa enzymatic activity is observed, suggesting that the beta-helical structure may be able to accommodate a portion of the prion protein sequence and, as a corollary, that a prion protein fragment may adopt left-handed parallel beta helix (LbetaH) architecture
additional information
-
a recombinant prion protein-LpxA protein is generated, in which a PrP fragment that is thought to be essential for the conformational conversion is incorporated into the beta-helical domain of LpxA. Partial Lpxa enzymatic activity is observed, suggesting that the beta-helical structure may be able to accommodate a portion of the prion protein sequence and, as a corollary, that a prion protein fragment may adopt left-handed parallel beta helix (LbetaH) architecture
additional information
-
heterologous expression of Acidithiobacillus ferrooxidans' genes lpxA, gnnA, and gnnB results in occurence of UDP-2-acetamido-3-amino-2,3-dideoxy-alpha-D-glucopyranose utilizing activity in Escherichia coli cells with defective lpxA gene in vivo
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
using Ni-NTA chromatography and gel filtration. Around 15 mg of purified protein is obtained from 1 l of bacterial culture
recombinant LpxA from Echerichia coli strain BL21-AI/pET24a by Reactive Green 19 affinity chromatography, dialysis, and ion exchange chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene lpxA, cloning from strain strain MG1655, subcloning in strain XL-1 Blue, expression in Escherichia coli strain BL21-AI/pET24a
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
drug development
the enzyme is a target for design of inhibitors with antibiotic potency
medicine
structures helps to design new inhibitors that target LpxA, antibiotics
drug development
drug development
-
development of small molecule, dual-binding LpxA/LpxD inhibitors as novel antimicrobials
drug development
-
LpxA is a potential target for antibacterial drug discovery
drug development
-
Lpxa is a viable target for antimicrobial drug development
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Anderson, M.S.; Bulawa, C.E.; Raetz, C.R.
The biosynthesis of gram-negative endotoxin. Formation of lipid A precursors from UDP-GlcNAc in extracts of Escherichia coli
J. Biol. Chem.
260
15536-15541
1985
Escherichia coli, Escherichia coli JB1104
Anderson, M.S.; Raetz, C.R.
UDP-N-acetylglucosamine 3-O-acyltransferase from Escherichia coli
Methods Enzymol.
209
449-454
1992
Escherichia coli
Anderson, M.S.; Raetz, C.R.
Biosynthesis of lipid A precursors in Escherichia coli. A cytoplasmic acyltransferase that converts UDP-N-acetylglucosamine to UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine
J. Biol. Chem.
262
5159-5169
1987
Escherichia coli, Escherichia coli JB1104, Escherichia coli W3106
Anderson, M.S.; Bull, H.G.; Galloway, S.M.; Kelly, T.M.; Mohan, S.; Radika, K.; Raetz, C.R.
UDP-N-acetylglucosamine acyltransferase of Escherichia coli. The first step of endotoxin biosynthesis is thermodynamically unfavorable
J. Biol. Chem.
268
19858-19865
1993
Escherichia coli
Williamson, J.M.; Anderson, M.S.; Raetz, C.R.
Acyl-acyl carrier protein specificity of UDP-GlcNAc acyltransferases from gram-negative bacteria: relationship to lipid A structure
J. Bacteriol.
173
3591-3596
1991
Acinetobacter calcoaceticus, Cereibacter sphaeroides, Citrobacter freundii, Escherichia coli, Escherichia coli JB1104, Escherichia coli SM105, Klebsiella aerogenes, Klebsiella oxytoca, Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens
Vaara, M.
Eight bacterial proteins, including UDP-N-acetylglucosamine acyltransferase (LpxA) and three other transferases of Escherichia coli, consist of a six-residue periodicity theme
FEMS Microbiol. Lett.
97
249-254
1992
Escherichia coli
-
Raetz, C.R.H.; Roderick, S.L.
A left-handed parallel beta helix in the structure of UDP-N-acetylglucosamine acyltransferase
Science
270
997-1000
1995
Escherichia coli
Sorensen, P.G.; Lutkenhaus, J.; Young, K.; Eveland, S.S.; Anderson, M.S.; Raetz, C.R.H.
Regulation of UDP-3-O-[R-3-hydroxymyristoyl]-N-acetylglucosamine deacetylase in Escherichia coli. The second enzymic step of lipid A biosynthesis
J. Biol. Chem.
271
25898-25905
1996
Escherichia coli, Escherichia coli SM105
Wyckoff, T.J.O.; Raetz, C.R.H.
The active site of Escherichia coli UDP-N-acetylglucosamine acyltransferase. Chemical modification and site-directed mutagenesis
J. Biol. Chem.
274
27047-27055
1999
Escherichia coli
Odegaard, T.J.; Kaltashov, I.A.; Cotter, R.J.; Steeghs, L.; Van Der Ley, P.; Khan, S.; Maskell, D.J.; Raetz, C.R.H.
Shortened hydroxyacyl chains on lipid A of Escherichia coli cells expressing a foreign UDP-N-acetylglucosamine O-acyltransferase
J. Biol. Chem.
272
19688-19696
1997
Escherichia coli, Neisseria meningitidis
Wyckoff, T.J.O.; Lin, S.; Cotter, R.J.; Dotson, G.D.; Raetz, C.R.H.
Hydrocarbon rulers in UDP-N-acetylglucosamine acyltransferases
J. Biol. Chem.
273
32369-32372
1998
Escherichia coli, Pseudomonas aeruginosa
Sweet, C.R.; Williams, A.H.; Karbarz, M.J.; Werts, C.; Kalb, S.R.; Cotter, R.J.; Raetz, C.R.
Enzymatic synthesis of lipid A molecules with four amide-linked acyl chains. LpxA acyltransferases selective for an analog of UDP-N-acetylglucosamine in which an amine replaces the 3"-hydroxyl group
J. Biol. Chem.
279
25411-25419
2004
Acidithiobacillus ferrooxidans (Q6QHI7), Escherichia coli, Leptospira interrogans, Mesorhizobium loti
Ulaganathan, V.; Buetow, L.; Hunter, W.N.
Nucleotide substrate recognition by UDP-N-acetylglucosamine acyltransferase (LpxA) in the first step of lipid A biosynthesis
J. Mol. Biol.
369
305-312
2007
Escherichia coli (P0A722), Escherichia coli
Williams, A.H.; Immormino, R.M.; Gewirth, D.T.; Raetz, C.R.
Structure of UDP-N-acetylglucosamine acyltransferase with a bound antibacterial pentadecapeptide
Proc. Natl. Acad. Sci. USA
103
10877-10882
2006
Escherichia coli (P0A722), Escherichia coli
Williams, A.H.; Raetz, C.R.
Structural basis for the acyl chain selectivity and mechanism of UDP-N-acetylglucosamine acyltransferase
Proc. Natl. Acad. Sci. USA
104
13543-13550
2007
Escherichia coli (P0A722), Escherichia coli
Gong, H.; Murphy, P.W.; Langille, G.M.; Minielly, S.J.; Murphy, A.; McMaster, C.R.; Byers, D.M.
Tryptophan fluorescence reveals induced folding of Vibrio harveyi acyl carrier protein upon interaction with partner enzymes
Biochim. Biophys. Acta
1784
1835-1843
2008
Escherichia coli
Choi, J.H.; May, B.C.; Govaerts, C.; Cohen, F.E.
Site-directed mutagenesis demonstrates the plasticity of the beta helix: implications for the structure of the misfolded prion protein
Structure
17
1014-1023
2009
Escherichia coli (P0A722), Escherichia coli
Jenkins, R.J.; Dotson, G.D.
Dual targeting antibacterial peptide inhibitor of early lipid a biosynthesis
ACS Chem. Biol.
7
1170-1177
2012
Escherichia coli
Jenkins, R.J.; Dotson, G.D.
A continuous fluorescent enzyme assay for early steps of lipid A biosynthesis
Anal. Biochem.
425
21-27
2012
Escherichia coli
Das, A.; Mukhopadhyay, C.
LpxA: a natural nanotube
Biopolymers
93
845-853
2010
Escherichia coli
Shapiro, A.B.; Ross, P.L.; Gao, N.; Livchak, S.; Kern, G.; Yang, W.; Andrews, B.; Thresher, J.
A high-throughput-compatible fluorescence anisotropy-based assay for competitive inhibitors of Escherichia coli UDP-N-acetylglucosamine acyltransferase (LpxA)
J. Biomol. Screen.
18
341-347
2013
Escherichia coli
Jenkins, R.J.; Heslip, K.A.; Meagher, J.L.; Stuckey, J.A.; Dotson, G.D.
Structural basis for the recognition of peptide RJPXD33 by acyltransferases in lipid A biosynthesis
J. Biol. Chem.
289
15527-15535
2014
Escherichia coli (P0A722), Escherichia coli
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