Literature summary for 4.1.3.38 extracted from

ORourke, P.E.; Eadsforth, T.C.; Fyfe, P.K.; Shepherd, S.M.; Hunter, W.N.
Pseudomonas aeruginosa 4-amino-4-deoxychorismate lyase: spatial conservation of an active site tyrosine and classification of two types of enzyme (2011), PLoS ONE, 6, e24158.

Application

Application	Comment	Organism
drug development	the absence of the enzyme in humans and its essentiality in various microbes suggests that inhibition of PabC offers the possibility of therapeutics targeting a range of microbial infections, potential of this protein for early stage drug discovery	Pseudomonas aeruginosa

Cloned(Commentary)

Cloned (Comment)	Organism
gene pabC, expression of His6-tagged enzyme with TEV protease cleavage site in Escherichia coli strain BL21(DE3) GOLD	Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant detagged enzyme, hanging drop vapour diffusion method, mixing of 0.001 ml of 33 mg/mL prrotein in 100 mM HEPES, pH 7.5, 500 mM NaCl, 0.1 mM pyridoxal 5'-phosphate, and 10 mM 4-aminobenzoate, with 0001 ml of reservoir solution containing 10% w/v PEG 400, 1.8 M ammonium sulfate and 100 mM MES, pH 6.5, 20°C, 1 week, X-ray diffraction structure dtermination and analysis at 1.75 A resolution	Pseudomonas aeruginosa

Crystallization (Comment)

Organism

purified recombinant detagged enzyme, hanging drop vapour diffusion method, mixing of 0.001 ml of 33 mg/mL prrotein in 100 mM HEPES, pH 7.5, 500 mM NaCl, 0.1 mM pyridoxal 5'-phosphate, and 10 mM 4-aminobenzoate, with 0001 ml of reservoir solution containing 10% w/v PEG 400, 1.8 M ammonium sulfate and 100 mM MES, pH 6.5, 20°C, 1 week, X-ray diffraction structure dtermination and analysis at 1.75 A resolution

Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
4-amino-4-deoxychorismate	Pseudomonas aeruginosa	-	4-aminobenzoate + pyruvate	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas aeruginosa	Q9HZN6	gene pabC	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) GOLD by nickel affinity chromatography, cleavage of the tag by TEV protease, and gel filtration	Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-amino-4-deoxychorismate	-	Pseudomonas aeruginosa	4-aminobenzoate + pyruvate	-	?
4-amino-4-deoxychorismate	molecular modeling of the catalytic intermediate, overview	Pseudomonas aeruginosa	4-aminobenzoate + pyruvate	-	?

Subunits

Subunits	Comment	Organism
dimer	sequence comparisons, structure modeling of the catalytic intermediate and ligand-bound enzyme, overview	Pseudomonas aeruginosa

Synonyms

Synonyms	Comment	Organism
4-amino-4-deoxychorismate lyase	-	Pseudomonas aeruginosa
PabC	-	Pseudomonas aeruginosa

Cofactor

Cofactor	Comment	Organism	Structure
pyridoxal 5'-phosphate	dependent on, the PLP methyl group makes van der Waals interactions with Gln147 and is positioned 3.2 A distant from the carbonyl oxygen of Val175, Ser237 interacts with the pyridoxal 5'-phosphate phosphate	Pseudomonas aeruginosa

General Information

General Information	Comment	Organism
evolution	structure comparisons with related enzymes, overview. PabC enzymes can be classified into two groups depending upon whether an active site and structurally conserved tyrosine is provided from the polypeptide that mainly forms an active site or from the partner subunit in the dimeric assembly	Pseudomonas aeruginosa
additional information	structure-activity relationship of PabC, ligand binding modeling and reaction mechanism, overview. No structure of PabC in complex with ligands is achieved, but a computational model of the catalytic intermediate docked into the enzyme active site is generated. A conserved tyrosine helps to create a hydrophobic wall on one side of the active site that provides important interactions to bind the catalytic intermediate, but it does not appear to participate in interactions with the C atom that undergoes an sp2 to sp3 conversion as pyruvate is produced. An active site threonine hydroxyl contributes a proton used in the reduction of the substrate methylene to pyruvate methyl in the final stage of the mechanism	Pseudomonas aeruginosa
physiological function	the enzyme is active in folate biosynthesis and essential for the cell growth of the pathogen	Pseudomonas aeruginosa