Literature summary for 2.5.1.54 extracted from

Cross, P.J.; Heyes, L.C.; Zhang, S.; Nazmi, A.R.; Parker, E.J.
The functional unit of Neisseria meningitidis 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase is dimeric (2016), PLoS ONE, 11, e0145187.

Cloned(Commentary)

Cloned (Comment)	Organism
gene aroG, recombinant expression of mutant enzyme R126S in Escherichia coli strain BL21 StarTM (DE3)	Neisseria meningitidis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme mutant R126S, hanging drop vapour diffusion method, mixing of 0.001 ml of 10 mg/mL protein solution with 0.001 ml of crystallisation buffer containing 0.1 Tris HCl, pH 7.3, 0.2 M trimethyl-amino-N-oxide, 0.6 mM MnSO4, and 15-20% w/v PEG 2000MME, equilibration over 0.5 ml of reservoir solution, 20°C, 7 days, X-ray diffraction structure determination and analysis at 2.0 A resolution	Neisseria meningitidis

Crystallization (Comment)

Organism

purified recombinant enzyme mutant R126S, hanging drop vapour diffusion method, mixing of 0.001 ml of 10 mg/mL protein solution with 0.001 ml of crystallisation buffer containing 0.1 Tris HCl, pH 7.3, 0.2 M trimethyl-amino-N-oxide, 0.6 mM MnSO4, and 15-20% w/v PEG 2000MME, equilibration over 0.5 ml of reservoir solution, 20°C, 7 days, X-ray diffraction structure determination and analysis at 2.0 A resolution

Neisseria meningitidis

Protein Variants

Protein Variants	Comment	Organism
R126S	site-directed mutagenesis, mutation of the residue involved in the salt bridge formation of Arg126-Glu27 results in perturbation of the less extensive interface in the enzyme tetramer and formation of enzyme dimers in solution. The dimeric NmeDAH7PSR126S variant exhibits a slight reduction in thermal stability by differential scanning calorimetry experiments and a slow loss of activity over time compared to the wild-type enzyme. Although NmeDAH7PSR126S crystallised as a tetramer, like the wild-type enzyme, structural asymmetry at the less extensive interface was observed consistent with its destabilisation	Neisseria meningitidis

Inhibitors

Inhibitors	Comment	Organism
L-phenylalanine	allosteric feedback inhibition	Neisseria meningitidis
L-tryptophan	allosteric feedback inhibition	Neisseria meningitidis
L-tyrosine	allosteric feedback inhibition	Neisseria meningitidis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0134	-	D-erythrose 4-phosphate	pH 6.8, 25°C, mutant R126S	Neisseria meningitidis
0.015	-	phosphoenolpyruvate	pH 6.8, 25°C, wild-type enzyme	Neisseria meningitidis
0.025	-	phosphoenolpyruvate	pH 6.8, 25°C, mutant R126S	Neisseria meningitidis
0.037	-	D-erythrose 4-phosphate	pH 6.8, 25°C, wild-type enzyme	Neisseria meningitidis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
38700	-	-	Neisseria meningitidis
66000	-	homodimeric mutant R126S enzyme, gel filtration	Neisseria meningitidis
69000	-	homodimeric mutant R126S enzyme, analytical ultracentrifugation	Neisseria meningitidis
141000	-	homotetrameric wild-type enzyme, analytical ultracentrifugation	Neisseria meningitidis
150000	-	homotetrameric wild-type enzyme, gel filtration	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	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant mutant enzyme R126S from Escherichia coli strain BL21 StarTM (DE3)	Neisseria meningitidis

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	-	?

Subunits

Subunits	Comment	Organism
homotetramer	4 * 38700, about, sequence calculation	Neisseria meningitidis
More	the enzyme forms a homotetrameric structure with an extensive and a less extensive interface, but the functional unit of the enzyme is dimeric. The tetrameric association enabled by this Arg126-Glu27 salt-bridge appears to contribute solely to the stability of the protein. Analysis of quarternary structure, PDB ID 4HSN, and SAXS data analysis	Neisseria meningitidis

Synonyms

Synonyms	Comment	Organism
3-deoxy-D-arabino-heptulosonate 7-phosphate synthase	-	Neisseria meningitidis
Nme-DAH7PS	-	Neisseria meningitidis

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
12.6	-	phosphoenolpyruvate	pH 6.8, 25°C, mutant R126S	Neisseria meningitidis
12.6	-	D-erythrose 4-phosphate	pH 6.8, 25°C, mutant R126S	Neisseria meningitidis
27.1	-	phosphoenolpyruvate	pH 6.8, 25°C, wild-type enzyme	Neisseria meningitidis
27.1	-	D-erythrose 4-phosphate	pH 6.8, 25°C, wild-type enzyme	Neisseria meningitidis

pH Optimum

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

General Information

General Information	Comment	Organism
evolution	despite significant sequence diversity, all known DAH7PS enzymes share a common (beta/alpha)8-barrel core and similar active site architectures formed by conserved residues that support a similar catalytic mechanism. On the basis of sequence and size, DAH7PS enzymes are classified into groups (type Ialpha, type Ibeta and type II). Each group features distinct structural elements appended to the core catalytic barrel that are associated with the allosteric inhibition of the enzymes and different quaternary structure associations. Neisseria meningitidis DAH7PS belongs to the type Ialpha group	Neisseria meningitidis
metabolism	3-deoxy-D-arabino-heptulosonate 7-phosphate synthase catalyses the first committed step of the shikimate pathway, which involves the aldol-like condensation of D-erythrose 4-phosphate and phosphoenolpyruvate to form 3-deoxy-D-arabino-heptulosonate 7-phosphate	Neisseria meningitidis
physiological function	3-deoxy-D-arabino-heptulosonate 7-phosphate synthase catalyses the aldol-like condensation of D-erythrose 4-phosphate and phosphoenolpyruvate to form 3-deoxy-D-arabino-heptulosonate 7-phosphate. Entry into the pathway is controlled by the allosteric regulation of enzyme DAH7PS by the pathway end products Phe, Tyr and Trp, or other intermediates of the pathway	Neisseria meningitidis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
504	-	phosphoenolpyruvate	pH 6.8, 25°C, mutant R126S	Neisseria meningitidis
732	-	D-erythrose 4-phosphate	pH 6.8, 25°C, wild-type enzyme	Neisseria meningitidis
940	-	D-erythrose 4-phosphate	pH 6.8, 25°C, mutant R126S	Neisseria meningitidis
1806	-	phosphoenolpyruvate	pH 6.8, 25°C, wild-type enzyme	Neisseria meningitidis