Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Bartling, C.M.; Raetz, C.R.
    Steady-state kinetics and mechanism of LpxD, the N-acyltransferase of lipid A biosynthesis (2008), Biochemistry, 47, 5290-5302.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.3.1.191 additional information LpxD does not require the presence of a detergent for catalytic activity because the critical micelle concentrations of its substrates are likely to be above 0.1 mM Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.191 overexpression in Escherichia coli Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.3.1.191 D232A mutation causes a 10fold reduction in kcat and a striking increase in the KM for both substrates Escherichia coli
2.3.1.191 F41A mutation increases the KM for UDP-3-O-((3R)-3-hydroxymyristoyl)-a-D-glucosamine 30fold and kcat 5fold Escherichia coli
2.3.1.191 K194A mutation has little effect on activity Escherichia coli
2.3.1.191 K46A mutation causes 3fold increase in KM((3R)-3-hydroxymyristoyl-[acyl-carrier protein]) and has no effect on kcat Escherichia coli
2.3.1.191 N233A mutation causes a 10fold reduction in kcat and a striking increase in the KM for both substrates Escherichia coli
2.3.1.191 N240A causes less than a 2fold reduction in specific activity, when assayed at substrate concentrations at 2fold above KM with the purified proteins Escherichia coli
2.3.1.191 N44A causes less than a 2fold reduction in specific activity, when assayed at substrate concentrations at 2fold above KM with the purified proteins Escherichia coli
2.3.1.191 Q236A mutation has little effect on activity Escherichia coli
2.3.1.191 Q32A causes less than a 2fold reduction in specific activity, when assayed at substrate concentrations at 2fold above KM with the purified proteins Escherichia coli
2.3.1.191 R293A KM((3R)-3-hydroxymyristoyl-[acyl-carrier protein]) increases 23fold compared to wild-type with little effect on kcat Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.1.191 (3R)-3-hydroxylauroyl-methylphosphopantetheine competitive inhibitor with respect to UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine and an uncompetitive inhibitor with respect to (3R)-3-hydroxymyristoyl-[acyl-carrier protein] Escherichia coli
2.3.1.191 (3R)-3-hydroxylauroylmethylphosphopantetheine uncompetitive inhibitor against (3R)-3-hydroxymyristoyl-[acyl-carrier protein] and a competitive inhibitor against UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine Escherichia coli
2.3.1.191 (3R)-3-hydroxymyristoyl-[acyl-carrier protein]
-
 Escherichia coli
2.3.1.191 acyl-carrier protein competitive inhibitor with respect to (3R)-3-hydroxymyristoyl-[acyl-carrier protein] and a noncompetitive inhibitor with respect to UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine Escherichia coli
2.3.1.191 Ca2+ inhibition is overcome by the addition of excess (3R)-3-hydroxymyristoyl-[acyl-carrier protein] Escherichia coli
2.3.1.191 Mg2+
-
 Escherichia coli
2.3.1.191 additional information divalent cations inhibit (3R)-3-hydroxymyristoyl-[acyl-carrier protein]-dependent acylation but not (3R)-3-hydroxylauroylmethylphosphopantetheine-dependent acylation, indicating that the acidic recognition helix of (3R)-3-hydroxymyristoyl-[acyl-carrier protein] contributes to binding; Na+ and K+ ions do not inhibit LpxD activity Escherichia coli
2.3.1.191 UDP-2-N-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine noncompetitive inhibitor against both substrates Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.191 additional information
-
 additional information steady-state kinetic analysis Escherichia coli
2.3.1.191 0.00084
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme H239A Escherichia coli
2.3.1.191 0.0017
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme H276A Escherichia coli
2.3.1.191 0.0032
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, wild-type enzyme Escherichia coli
2.3.1.191 0.0034
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme H239A Escherichia coli
2.3.1.191 0.0035
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme F41A Escherichia coli
2.3.1.191 0.0036
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme R293A Escherichia coli
2.3.1.191 0.0038
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme K194A Escherichia coli
2.3.1.191 0.0042
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme Q236A Escherichia coli
2.3.1.191 0.005
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme Y47A Escherichia coli
2.3.1.191 0.0056
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme K194A Escherichia coli
2.3.1.191 0.0063
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme H276A Escherichia coli
2.3.1.191 0.0071
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme K46A Escherichia coli
2.3.1.191 0.0078
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme Y47A Escherichia coli
2.3.1.191 0.012
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme K46A Escherichia coli
2.3.1.191 0.013
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme N233A Escherichia coli
2.3.1.191 0.013
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme Q236A Escherichia coli
2.3.1.191 0.028
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme N233A Escherichia coli
2.3.1.191 0.047
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme D232A Escherichia coli
2.3.1.191 0.067
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme D232A Escherichia coli
2.3.1.191 0.073
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme F41A Escherichia coli
2.3.1.191 0.074
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme R293A Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.3.1.191 35880
-
 3 * 35880, electrospray-ionization/time-of-flight mass spectrometry Escherichia coli
2.3.1.191 35881
-
 3 * 35881, calculated from sequence Escherichia coli
2.3.1.191 108000
-
 gel filtration Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.191 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine Escherichia coli third step of lipid A biosynthesis UDP-2,3-bis(3-hydroxymyristoyl)-alpha-D-glucosamine + holo-[acyl-carrier protein]
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.191 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.191 purification of untagged EcLpxD and an active N-terminally His6-tagged LpxD variant to near homogeneityrecombinant enzyme Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.1.191 7.14
-
 specific activity in the absence of Ca2+ Escherichia coli
2.3.1.191 8.9
-
-
 Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.191 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine third step of lipid A biosynthesis Escherichia coli UDP-2,3-bis(3-hydroxymyristoyl)-alpha-D-glucosamine + holo-[acyl-carrier protein]
-
 ?
2.3.1.191 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] + UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine compulsory ordered mechanism in which (3R)-3-hydroxymyristoyl-[acyl-carrier protein] binds prior to UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine. The product, UDP-2,3-diacylglucosamine, dissociates prior to acyl-carrier protein Escherichia coli UDP-2,3-bis(3-hydroxymyristoyl)-alpha-D-glucosamine + holo-[acyl-carrier protein]
-
 ?
2.3.1.191 additional information R-3-hydroxylauroyl-methylphosphopantetheine is a very poor substrate. The specific activity, measured at either 0.01 mM or 1 mM (3R)-3-hydroxylauroylmethylphosphopantetheine as the acyl donor, is more than 100fold lower than with 0.01 mM (3R)-3-hydroxymyristoyl-[acyl-carrier protein] Escherichia coli ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.1.191 homotrimer 3 * 35880, electrospray-ionization/time-of-flight mass spectrometry Escherichia coli
2.3.1.191 homotrimer 3 * 35881, calculated from sequence Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
2.3.1.191 EcLpxD
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.191 30
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.191 additional information
-
 additional information steady-state kinetic analysis Escherichia coli
2.3.1.191 0.032
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme H239A Escherichia coli
2.3.1.191 0.032
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme H239A Escherichia coli
2.3.1.191 0.73
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme H276A Escherichia coli
2.3.1.191 0.73
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme H276A Escherichia coli
2.3.1.191 1.4
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme N233A Escherichia coli
2.3.1.191 1.4
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme N233A Escherichia coli
2.3.1.191 1.9
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme D232A Escherichia coli
2.3.1.191 1.9
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme D232A Escherichia coli
2.3.1.191 4.1
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme F41A Escherichia coli
2.3.1.191 4.1
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme F41A Escherichia coli
2.3.1.191 5.7
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme K194A Escherichia coli
2.3.1.191 5.7
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme K194A Escherichia coli
2.3.1.191 8.9
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme R293A Escherichia coli
2.3.1.191 8.9
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme R293A Escherichia coli
2.3.1.191 12
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme Q236A Escherichia coli
2.3.1.191 12
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme Q236A Escherichia coli
2.3.1.191 17
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme K46A Escherichia coli
2.3.1.191 17
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme K46A Escherichia coli
2.3.1.191 18
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme Y47A Escherichia coli
2.3.1.191 18
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, mutant enzyme Y47A Escherichia coli
2.3.1.191 23
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, wild-type enzyme Escherichia coli
2.3.1.191 23
-
 UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, wild-type enzyme Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.191 7.5
-
 assay at Escherichia coli

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3.1.191 0.0043
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C Escherichia coli
2.3.1.191 0.006
-
 UDP-2-N-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, noncompetitive inhibition against (R)-3-hydroxymyristoyl-[acyl-carrier protein] Escherichia coli
2.3.1.191 0.0094
-
 UDP-2-N-(R-3-hydroxymyristoyl)-alpha-D-glucosamine pH 7.5, 30°C, noncompetitive inhibition against UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine Escherichia coli
2.3.1.191 0.048
-
 acyl-carrier protein pH 7.5, 30°C, competitive inhibition with respect to (R)-3-hydroxymyristoyl-[acyl-carrier protein] Escherichia coli
2.3.1.191 0.139
-
 acyl-carrier protein pH 7.5, 30°C, noncompetitive inhibitionr with respect to UDP-3-O-(R-3-hydroxymyristoyl)-alpha-D-glucosamine Escherichia coli
2.3.1.191 0.39
-
 (3R)-3-hydroxylauroyl-methylphosphopantetheine pH 7.5, 30°C, competitive inhibition with respect to UDP-3-O-((3R)-3-hydroxymyristoyl)-alpha-D-glucosamine Escherichia coli
2.3.1.191 0.69
-
 (3R)-3-hydroxylauroyl-methylphosphopantetheine pH 7.5, 30°C, uncompetitive inhibition with respect to (3R)-3-hydroxymyristoyl-[acyl-carrier protein] Escherichia coli

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.3.1.191 0.42
-
 pH 7.5, 30°C Escherichia coli Ca2+
2.3.1.191 1.41
-
 pH 7.5, 30°C Escherichia coli Mg2+

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.3.1.191 9.4
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme H239A Escherichia coli
2.3.1.191 10.7
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme N233A Escherichia coli
2.3.1.191 40.4
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme D232A Escherichia coli
2.3.1.191 120.3
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme R293A Escherichia coli
2.3.1.191 429
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme H276A Escherichia coli
2.3.1.191 923
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme Q236A Escherichia coli
2.3.1.191 1171
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme F41A Escherichia coli
2.3.1.191 1417
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme K46A Escherichia coli
2.3.1.191 1500
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme K194A Escherichia coli
2.3.1.191 3600
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, mutant enzyme Y47A Escherichia coli
2.3.1.191 7188
-
 (3R)-3-hydroxymyristoyl-[acyl-carrier protein] pH 7.5, 30°C, wild-type enzyme Escherichia coli