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Literature summary extracted from

  • Sasso, S.; Okvist, M.; Roderer, K.; Gamper, M.; Codoni, G.; Krengel, U.; Kast, P.
    Structure and function of a complex between chorismate mutase and DAHP synthase: efficiency boost for the junior partner (2009), EMBO J., 28, 2128-2142.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
5.4.99.5 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase the catalytic efficiency of chorismate mutase increases 140fold on addition of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase, chorismate mutase forms a complex with 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase and that complex formation both increases the CM activity by more than two orders of magnitude and endows chorismate mutase with regulatory features Mycobacterium tuberculosis
5.4.99.5 additional information neither pH variation between 5.9 and 8.7, nor provision of 0.1 mg/ml bovine serum albumin, 2 mM Ca2+, 10 mM Mg2+, 1 mM EDTA, 1 mM EGTA, 1 mM 1,10-phenanthroline, 1 mM L-phenylalanine, 1 mM L-tyrosine, 1 mM L-tryptophan, or 0.6 mM salicylate affect catalytic activity by more than a factor of 2 Mycobacterium tuberculosis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.54 N-terminally histidine-tagged enzyme is expressed in Escherichia coli KA13 Mycobacterium tuberculosis
5.4.99.5 expressed in Escherichia coli strain KA13 Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.5.1.54 in complex with chorismate mutase, streak-seeding conditions in 20 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5, 150 mM NaCl, 0.5 mM TCEP [tris(2-carboxyethyl)phosphine hydrochloride], 0.2 mM phosphoenolpyruvate, crystallization by 0.9 M ammonium sulfate, 100 mM Tris, pH 7.9 to 8.0, and 1 to 5% PEG 400 Mycobacterium tuberculosis
5.4.99.5 in complex with L-malate, hanging drop vapor diffusion method, using 15% (w/v) PEG 1500 and 0.1 M of the L-malate-containing MMT buffer system pH 8.0-9.0 Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
5.4.99.5 G86A mutant shows reduced kcat and Km values compared to the wild type enzyme Mycobacterium tuberculosis
5.4.99.5 K39R inactive Mycobacterium tuberculosis
5.4.99.5 L88A inactive Mycobacterium tuberculosis
5.4.99.5 R46K inactive Mycobacterium tuberculosis
5.4.99.5 R87A inactive Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.54 phenylalanine inhibition of the synthase-chorismate mutase complex, based on mutase activity measurements at 30°C, 50 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5, 0.5 mM TCEP [tris(2-carboxyethyl)phosphine hydrochloride], 0.2 mM phosphoenolpyruvate, and 0.1 mM MnCl2 Mycobacterium tuberculosis
2.5.1.54 tyrosine inhibition of the synthase-chorismate mutase complex, based on mutase activity measurements at 30°C, 50 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5, 0.5 mM TCEP [tris(2-carboxyethyl)phosphine hydrochloride], 0.2 mM phosphoenolpyruvate, and 0.1 mM MnCl2 Mycobacterium tuberculosis
5.4.99.5 L-Phe 28% residual activity at 0.2 mM Mycobacterium tuberculosis
5.4.99.5 L-Trp 96% residual activity at 0.2 mM Mycobacterium tuberculosis
5.4.99.5 L-Tyr 63% residual activity at 0.2 mM Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.4.99.5 0.57
-
 chorismate mutant enzyme G86A Mycobacterium tuberculosis
5.4.99.5 1.14
-
 chorismate wild type enzyme Mycobacterium tuberculosis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
5.4.99.5 intracellular
-
 Mycobacterium tuberculosis 5622
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.54 51830
-
 ESI-MS Mycobacterium tuberculosis
2.5.1.54 51830
-
 expected mass by calculation Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.54 phosphoenolpyruvate + erythrose 4-phosphate + H2O Mycobacterium tuberculosis
-
 3-deoxy-D-arabino-hept-2-ulosonate 7-phosphate + phosphate
-
 ?
2.5.1.54 phosphoenolpyruvate + erythrose 4-phosphate + H2O Mycobacterium tuberculosis H37Rv
-
 3-deoxy-D-arabino-hept-2-ulosonate 7-phosphate + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.54 Mycobacterium tuberculosis O53512
-
-
2.5.1.54 Mycobacterium tuberculosis H37Rv O53512
-
-
5.4.99.5 Mycobacterium tuberculosis P9WIC1
-
-
5.4.99.5 Mycobacterium tuberculosis H37Rv P9WIC1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.54 Ni-NTA affinity chromatography followed by gel filtration, in 20 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5, 150 mM NaCl, 0.5 mM TCEP [tris(2-carboxyethyl)phosphine hydrochloride], 0.2 mM phosphoenolpyruvate, and 0.1 mM MnCl2, ultrafiltration for concentration of enzyme Mycobacterium tuberculosis
5.4.99.5 anion exchange column chromatography, Superdex 75 gel filtration Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.54 phosphoenolpyruvate + erythrose 4-phosphate + H2O
-
 Mycobacterium tuberculosis 3-deoxy-D-arabino-hept-2-ulosonate 7-phosphate + phosphate
-
 ?
2.5.1.54 phosphoenolpyruvate + erythrose 4-phosphate + H2O
-
 Mycobacterium tuberculosis H37Rv 3-deoxy-D-arabino-hept-2-ulosonate 7-phosphate + phosphate
-
 ?
5.4.99.5 Chorismate
-
 Mycobacterium tuberculosis Prephenate
-
 ?
5.4.99.5 Chorismate
-
 Mycobacterium tuberculosis H37Rv Prephenate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.54 3-deoxy-7-phosphoheptulonate synthase
-
 Mycobacterium tuberculosis
2.5.1.54 DAHP synthase
-
 Mycobacterium tuberculosis
2.5.1.54 MtDS
-
 Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.4.99.5 0.11
-
 chorismate mutant enzyme G86A Mycobacterium tuberculosis
5.4.99.5 2
-
 chorismate wild type enzyme Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
2.5.1.54 metabolism shikimate biosynthetic pathway, noncovalent binding to chorismate mutase enhances the mutase activity more than 100fold, 4 enzyme molecules sandwiched between the chorismate mutase dimers reposition the active site of the mutases Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.54 84
-
 phosphoenolpyruvate results within factor two of this otherwise published value, 30°C, 50 mM BTP (1,3-bis[tris(hydroxymethyl)methylamino]propane), pH 7.5 with 100 microM MnCl2, 240 microM D-erythrose-4-phosphate (saturating), less than 10 microM phosphoenolpyruvate, no influence of chorismate mutase concentration on activity Mycobacterium tuberculosis
5.4.99.5 0.19
-
 chorismate mutant enzyme G86A Mycobacterium tuberculosis
5.4.99.5 1.75
-
 chorismate wild type enzyme Mycobacterium tuberculosis