Literature summary extracted from
Baum, B.; Lecker, L.S.; Zoltner, M.; Jaenicke, E.; Schnell, R.; Hunter, W.N.; Brenk, R.
Structures of Pseudomonas aeruginosa beta-ketoacyl-(acyl-carrier-protein) synthase II (FabF) and a C164Q mutant provide templates for antibacterial drug discovery and identify a buried potassium ion and a ligand-binding site that is an artefact of the crysta (2015), Acta Crystallogr. Sect. F, 71, 1020-1026.
Application
EC Number |
Application |
Comment |
Organism |
---|
2.3.1.179 |
drug development |
the enzyme is an antibiotic target in Gram-positive bacteria and might also be a target in Gram-negative bacteria |
Pseudomonas aeruginosa |
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.3.1.179 |
gene fabF, recombinant wild-type and mutant N-terminally His6-tagged enzyme expressions from vector pNIC28-Bsa4 including a TEV cleavage site in Escherichia coli strain BL21(DE3)pLysS |
Pseudomonas aeruginosa |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
2.3.1.179 |
purified recombinant free wild-type enzyme, and mutant C164Q enzyme, free or in complex with 3-(benzoylamino)-2-hydroxybenzoic acid, mixing of 0.001 ml of 20 mg/ml protein in 25 mM Tris-HCl, pH 7.5, 150 mM NaCl, with 0.001 ml reservoir solution containing 0.2 M MgCl2, 0.1 M Tris-HCl, pH 7.0, 10% w/v PEG 8000 for the wild-type enzyme, or 0.001 ml of 20 mg/ml protein in 50 mM Na2HPO4, pH 7.8, 150 mM NaCl, 10% v/v glycerol, and 0.5 mM DTT with 0.001 ml of reservoir consisting of 0.2 M NH4HCO2, 25% w/v PEG 3350 for the mutant enzyme, equilibration against 0.06 ml reservoir solution, at 20°C, X-ray diffraction structure determination and analysis at 1.67-2.46 A resolution, molecular replacement |
Pseudomonas aeruginosa |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
2.3.1.179 |
C164Q |
site-directed mutagenesis, a mutant in which the binding site is altered to resemble the substrate-bound state |
Pseudomonas aeruginosa |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
2.3.1.179 |
3-(benzoylamino)-2-hydroxybenzoic acid |
binds outside the active site of the enzyme, binding structure with wild-type and C164Q mutant enzymes, overview. Access to the depths of the active site of the PaFabF apoenzyme is restricted by the conformations of Phe230 and Phe400. 3-(benzoylamino)-2-hydroxybenzoic acid/Mg2+ ion pair selectively binds into and perhaps contributes to the formation of a stable binding site on the surface of the enzyme distant from the active site, from which it is likely to be occluded by steric hindrance |
Pseudomonas aeruginosa |
|
2.3.1.179 |
platensimycin |
a natural product inhibitor |
Pseudomonas aeruginosa |
|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
2.3.1.179 |
55000 |
- |
recombinant enzyme, gel filtration |
Pseudomonas aeruginosa |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.3.1.179 |
Pseudomonas aeruginosa |
G3XDA2 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.3.1.179 |
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS by nickel affinit chromatography, tag cleavage through TEV, dialysis, and gel filtration |
Pseudomonas aeruginosa |
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
2.3.1.179 |
dimer |
- |
Pseudomonas aeruginosa |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
2.3.1.179 |
3-oxoacyl-ACP synthase II |
- |
Pseudomonas aeruginosa |
2.3.1.179 |
beta-ketoacyl-(acyl-carrier-protein) synthase II |
- |
Pseudomonas aeruginosa |
2.3.1.179 |
FabF |
- |
Pseudomonas aeruginosa |
General Information
EC Number |
General Information |
Comment |
Organism |
---|
2.3.1.179 |
metabolism |
the enzyme is involved in fatty-acid biosynthesis |
Pseudomonas aeruginosa |
2.3.1.179 |
additional information |
active site structure of wild-type and mutant enzymes, ligand binding structures, overview |
Pseudomonas aeruginosa |
2.3.1.179 |
physiological function |
the enzyme is involved in fatty-acid biosynthesis |
Pseudomonas aeruginosa |