Literature summary for 6.2.1.32 extracted from

Witzgall, F.; Ewert, W.; Blankenfeldt, W.
Structures of the N-terminal domain of PqsA in complex with anthraniloyl- and 6-fluoroanthraniloyl-AMP substrate activation in Pseudomonas quinolone signal (PQS) biosynthesis (2017), ChemBioChem, 18, 2045-2055 .

Application

Application	Comment	Organism
drug development	the enzyme might be an attractive target for anti-infective agents	Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified isolated N-terminal domain of anthranilate-CoA ligase PqsA in complex with anthraniloyl- and 6-fluoroanthraniloyl-AMP, sitting drop vapor diffusion technique, mixing of 200 nl of protein solution with 200 nl of reservoir solution containing 0.1 M Na3 citrate, pH 5.8-6.3, 22-28% w/v PEG 3350, and 0.2-0.5 M NH4OAc. Crystals can only be obtained in the presence of substrates, leading to complexes with anthraniloyl-AMP or 6-fluoroanthraniloyl-AMP, 20°C, X-ray diffraction structure determination and analysis at 1.43-1.90 A resolution, molecular replacement using the structure of the benzoate-CoA ligase from Bacillus xenoverans LB400, PDB ID 2V7B, as a search model, modeling	Pseudomonas aeruginosa

Crystallization (Comment)

Organism

purified isolated N-terminal domain of anthranilate-CoA ligase PqsA in complex with anthraniloyl- and 6-fluoroanthraniloyl-AMP, sitting drop vapor diffusion technique, mixing of 200 nl of protein solution with 200 nl of reservoir solution containing 0.1 M Na3 citrate, pH 5.8-6.3, 22-28% w/v PEG 3350, and 0.2-0.5 M NH4OAc. Crystals can only be obtained in the presence of substrates, leading to complexes with anthraniloyl-AMP or 6-fluoroanthraniloyl-AMP, 20°C, X-ray diffraction structure determination and analysis at 1.43-1.90 A resolution, molecular replacement using the structure of the benzoate-CoA ligase from Bacillus xenoverans LB400, PDB ID 2V7B, as a search model, modeling

Pseudomonas aeruginosa

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + anthranilate + CoA	Pseudomonas aeruginosa	-	AMP + diphosphate + anthraniloyl-CoA	-	?
ATP + anthranilate + CoA	Pseudomonas aeruginosa ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1	-	AMP + diphosphate + anthraniloyl-CoA	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas aeruginosa	Q9I4X3	-	-
Pseudomonas aeruginosa ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1	Q9I4X3	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
ATP + anthranilate + CoA = AMP + diphosphate + anthraniloyl-CoA	reaction via anthraniloyl-AMP intermediate. The catalytic cycle of PqsA consists of two steps: in the first half-reaction of PqsA (adenylation), anthranilate is activated with ATP/Mg2+ to produce the intermediate anthraniloyl-AMP with the release of diphosphate. The aryl moiety is then transferred to the acceptor molecule CoA to form anthraniloyl-CoA in the second half-reaction (thioester formation). The two half-reactions are cooperatively catalyzed by a large N-terminal domain and a smaller C-terminal domain separated by a flexible linker, the latter undergoes large conformational rearrangements in the course of the catalytic cycle	Pseudomonas aeruginosa	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + anthranilate + CoA	-	Pseudomonas aeruginosa	AMP + diphosphate + anthraniloyl-CoA	-	?
ATP + anthranilate + CoA	-	Pseudomonas aeruginosa ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1	AMP + diphosphate + anthraniloyl-CoA	-	?
additional information	6-fluoroanthraniloyl-AMP (6FABA-AMP), an inhibitor of PQS biosynthesis, is a good substrate of enzyme PqsA, enzyme binding structure analysis of 6-fluoroanthraniloyl-AMP and anthraniloyl-AMP, overview. 6FABA-AMP binds to PqsANTD in a similar way to anthraniloyl-AMP, by establishing water-mediated hydrogen bonds between the 2-amino group and Q162	Pseudomonas aeruginosa	?	-	?
additional information	6-fluoroanthraniloyl-AMP (6FABA-AMP), an inhibitor of PQS biosynthesis, is a good substrate of enzyme PqsA, enzyme binding structure analysis of 6-fluoroanthraniloyl-AMP and anthraniloyl-AMP, overview. 6FABA-AMP binds to PqsANTD in a similar way to anthraniloyl-AMP, by establishing water-mediated hydrogen bonds between the 2-amino group and Q162	Pseudomonas aeruginosa ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1	?	-	?

Subunits

Subunits	Comment	Organism
More	The two half-reactions are cooperatively catalyzed by a large N-terminal domain and a smaller C-terminal domain separated by a flexible linker, the latter undergoes large conformational rearrangements in the course of the catalytic cycle. In PqsA, the adenine ring of the complexed adenylate moiety is sandwiched between the main-chain atoms of G279, S280, and P281 from a loop between beta12 and alpha12 on one side and the side chain of I301 on the opposite side, structure analysis, overview	Pseudomonas aeruginosa

Synonyms

Synonyms	Comment	Organism
anthranilate-CoA ligase	-	Pseudomonas aeruginosa
PqsA	-	Pseudomonas aeruginosa

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Pseudomonas aeruginosa

General Information

General Information	Comment	Organism
evolution	PqsA belongs to the ANL superfamily of anthranilate-CoA ligases that recognize the amino group of anthranilate through a water-mediated hydrogen bond	Pseudomonas aeruginosa
metabolism	the enzyme performs substrate activation in Pseudomonas aeruginosa quinolone signal (PQS) biosynthesis	Pseudomonas aeruginosa
additional information	active site structure and structure comparisons, overview. In PqsANTD, the 2-amino group of the anthraniloyl moiety is hydrogen-bonded to a conserved water molecule, which is coordinated by the side chain of Q162 and the carbonyl oxygen atom of G307. Q162 is located directly adjacent to the P-loop	Pseudomonas aeruginosa
physiological function	the enzyme performs substrate activation in Pseudomonas aeruginosa quinolone signal (PQS) biosynthesis	Pseudomonas aeruginosa