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Literature summary for 2.3.1.179 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.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
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)

Cloned (Comment) Organism
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)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
C164Q site-directed mutagenesis, a mutant in which the binding site is altered to resemble the substrate-bound state Pseudomonas aeruginosa

Inhibitors

Inhibitors Comment Organism Structure
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
platensimycin a natural product inhibitor Pseudomonas aeruginosa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
recombinant enzyme, gel filtration Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa G3XDA2
-
-

Purification (Commentary)

Purification (Comment) Organism
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

Subunits Comment Organism
dimer
-
Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
3-oxoacyl-ACP synthase II
-
Pseudomonas aeruginosa
beta-ketoacyl-(acyl-carrier-protein) synthase II
-
Pseudomonas aeruginosa
FabF
-
Pseudomonas aeruginosa

General Information

General Information Comment Organism
metabolism the enzyme is involved in fatty-acid biosynthesis Pseudomonas aeruginosa
additional information active site structure of wild-type and mutant enzymes, ligand binding structures, overview Pseudomonas aeruginosa
physiological function the enzyme is involved in fatty-acid biosynthesis Pseudomonas aeruginosa