Literature summary for 2.5.1.54 extracted from

Heyes, L.C.; Reichau, S.; Cross, P.J.; Jameson, G.B.; Parker, E.J.
Structural analysis of substrate-mimicking inhibitors in complex with Neisseria meningitidis 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase - the importance of accommodating the active site water (2014), Bioorg. Chem., 57, 242-250.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme complexed with inhibitors (S)-phospholactate, (R)-phospholactate, and vinyl phosphonate, hanging drop vapour diffusion method, mixing of 0.001 ml of 10 mg/mL protein in 10 mM bis(tris(hydroxymethyl)methylamino)propane, pH 7.3, and 5 mM inhibitor, with 0.001 ml of crystallisation buffer containing 0.1 Tris HCl, pH 7.3, 0.2 M trimethyl-amino-N-oxide, 0.4 mM MnSO4, and 15-20% w/v PEG 2000MME, equilibration over 0.5 ml of reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.76-2.34 A resolution	Neisseria meningitidis

Crystallization (Comment)

Organism

purified recombinant enzyme complexed with inhibitors (S)-phospholactate, (R)-phospholactate, and vinyl phosphonate, hanging drop vapour diffusion method, mixing of 0.001 ml of 10 mg/mL protein in 10 mM bis(tris(hydroxymethyl)methylamino)propane, pH 7.3, and 5 mM inhibitor, with 0.001 ml of crystallisation buffer containing 0.1 Tris HCl, pH 7.3, 0.2 M trimethyl-amino-N-oxide, 0.4 mM MnSO4, and 15-20% w/v PEG 2000MME, equilibration over 0.5 ml of reservoir solution, 20°C, 3 days, X-ray diffraction structure determination and analysis at 1.76-2.34 A resolution

Neisseria meningitidis

Inhibitors

Inhibitors	Comment	Organism
(R)-phospholactate	-	Neisseria meningitidis
(S)-phospholactate	-	Neisseria meningitidis
additional information	binding structure determination and analysis, overview. The side chains of enzyme residues Lys188, Arg167 and Arg236 and the main chain peptide N of Ala166 that interact with the phosphate moiety of phosphoenolpyruvate form similar interactions with the phosphate or phosphonate functionalities of the three ligands, consistent with this portion of each inhibitor being able to mimic the phosphoenolpyruvate binding characteristics well. The tetrameric quaternary structures of enzyme NmeDAH7PS bound with (S)-phospholactate and vinyl phosphonate are identical to that of the isomorphous phosphoenolpyruvate-bound parent structure, PDB ID 4HSN. The increased inhibitor potency observed for the vinyl phosphonate over the (R)-phospholactate suggests that the active site is better prepared to accommodate a planar rather than tetrahedral intermediate	Neisseria meningitidis
vinyl phosphonate	-	Neisseria meningitidis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Neisseria meningitidis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mn2+	required, Mn2+ coordinates the PEP carboxylate functionality and a water molecule	Neisseria meningitidis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
phosphoenolpyruvate + D-erythrose 4-phosphate + H2O	Neisseria meningitidis	-	3-deoxy-D-arabino-hept-2-ulosonate 7-phospate + phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Neisseria meningitidis	Q9K169	gene aroG	-

Reaction

Reaction	Comment	Organism	Reaction ID
phosphoenolpyruvate + D-erythrose 4-phosphate + H2O = 3-deoxy-D-arabino-hept-2-ulosonate 7-phosphate + phosphate	proposed mechanism of catalysis of NmeDAH7PS showing key nucleophilic role for active site WAT1, overview	Neisseria meningitidis	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
phosphoenolpyruvate + D-erythrose 4-phosphate + H2O	-	Neisseria meningitidis	3-deoxy-D-arabino-hept-2-ulosonate 7-phospate + phosphate	-	?
phosphoenolpyruvate + D-erythrose 4-phosphate + H2O	phosphoenolpyruvate binding structure analysis	Neisseria meningitidis	3-deoxy-D-arabino-hept-2-ulosonate 7-phospate + phosphate	-	?

Synonyms

Synonyms	Comment	Organism
3-deoxy-D-arabino-heptulosonate 7-phosphate synthase	-	Neisseria meningitidis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Neisseria meningitidis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.8	-	assay at	Neisseria meningitidis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism
0.0039	-	vinyl phosphonate	pH 6.8, 25°C, recombinant enzyme	Neisseria meningitidis
0.099	-	(R)-phospholactate	pH 6.8, 25°C, recombinant enzyme	Neisseria meningitidis
0.36	-	(S)-phospholactate	pH 6.8, 25°C, recombinant enzyme	Neisseria meningitidis