EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
5.4.99.5 | cysteine | - |
Arabidopsis thaliana | |
5.4.99.5 | histidine | - |
Arabidopsis thaliana | |
5.4.99.5 | additional information | although AtCM2 contains the putative regulatory effector binding domain, phenylalanine, tyrosine, and tryptophan do not affect its activity | Arabidopsis thaliana | |
5.4.99.5 | tryptophan | - |
Arabidopsis thaliana | |
5.4.99.5 | tryptophan | activates about 3fold | Arabidopsis thaliana |
EC Number | Cloned (Comment) | Organism |
---|---|---|
5.4.99.5 | AtCM1, sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta II (DE3) | Arabidopsis thaliana |
5.4.99.5 | AtCM3, sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta II (DE3) | Arabidopsis thaliana |
5.4.99.5 | recombinant His6-tagged wild-type enzyme from Escherichia coli strain Rosetta II (DE3) by nickel affinity chromatography, tag cleavage with thrombin, dialysis, benzamidine/nickel affinity chromatography, and gel filtration | Arabidopsis thaliana |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
5.4.99.5 | purified recombinant detagged wild-type enzyme in complex with phenylalanine or tyrosine, hanging drop vapordiffusion method, mixing 0f 0.001 ml of 9 mg/ml protein in 25 mM HEPES, pH 7.5, and 100 mM NaCl, with 0.001 ml of reservoir solution containing 30% PEG 400, 0.1 M HEPES, pH 7.5, 0.2 M MgCl2, and 1 mM of either phenylalanine or tyrosine, X-ray diffraction structure determination and analysis at 2.30-2.45 A resolution, molecular replacement using yeast chorismate mutase structure, PDB ID 4CSM, as a search model | Arabidopsis thaliana |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
5.4.99.5 | D132G | site-directed mutagenesis, the mutant enzyme kinetically resembles isozyme AtCM1, it retains activation by tryptophan, although to a lesser extent than observed with wild-type AtCM3 | Arabidopsis thaliana |
5.4.99.5 | G149A | site-directed mutagenesis, the mutation eliminates the effector action of both phenylalanine and tyrosine | Arabidopsis thaliana |
5.4.99.5 | G149D | site-directed mutagenesis, the mutation eliminates the effector action of both phenylalanine and tyrosine | Arabidopsis thaliana |
5.4.99.5 | G213A | site-directed mutagenesis, mutation in effector binding site, the mutation eliminates the effect of aromatic amino acids on enzymatic activity | Arabidopsis thaliana |
5.4.99.5 | G213P | site-directed mutagenesis, mutation in effector binding site, the mutation eliminates the effect of aromatic amino acids on enzymatic activity | Arabidopsis thaliana |
5.4.99.5 | H145Q | site-directed mutagenesis, mutation in effector binding site, the mutation has varyring effects on the EC50 values for the aromatic amino acid effectors but does not change either positive or negative effects on enzymatic activity | Arabidopsis thaliana |
5.4.99.5 | additional information | generation of the expression construct N-terminally hexahistidine-tagged AtCM1 lacking the plastid localization peptide, AtCM1DELTA66 | Arabidopsis thaliana |
5.4.99.5 | R79k | site-directed mutagenesis, mutation in effector binding site, the mutation has varyring effects on the EC50 values for the aromatic amino acid effectors but does not change either positive or negative effects on enzymatic activity | Arabidopsis thaliana |
5.4.99.5 | V217T | site-directed mutagenesis, mutation in effector binding site | Arabidopsis thaliana |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
5.4.99.5 | additional information | although AtCM2 contains the putative regulatory effector binding domain, phenylalanine, tyrosine, and tryptophan do not affect its activity | Arabidopsis thaliana | |
5.4.99.5 | phenylalanine | model of the AtCM1x02phenylalanine complex including residues Arg79-Val290 and Val307-Asp340, the phenylalanine ligand, and 83 waters, inhibits about 20fold | Arabidopsis thaliana | |
5.4.99.5 | tyrosine | inhibits about 20fold | Arabidopsis thaliana |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.4.99.5 | additional information | - |
additional information | steady-state kinetics, isozyme AtCM1 shows Michaelis-Menten kinetics. Effect of aromatic amino acids on wild-type and mutant AtCM1, overview | Arabidopsis thaliana | |
5.4.99.5 | additional information | - |
additional information | steady-state kinetics, isozyme AtCM2, shows Michaelis-Menten kinetics | Arabidopsis thaliana | |
5.4.99.5 | additional information | - |
additional information | steady-state kinetics, isozyme AtCM3 displays positive cooperativity with a Hill coefficient of 2.1, indicating that substrate binding at one active site of the homodimer enhanced interaction at the second active site | Arabidopsis thaliana | |
5.4.99.5 | 0.15 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana | |
5.4.99.5 | 0.55 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana | |
5.4.99.5 | 1.1 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
5.4.99.5 | chloroplast | - |
Arabidopsis thaliana | 9507 | - |
5.4.99.5 | cytosol | - |
Arabidopsis thaliana | 5829 | - |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
5.4.99.5 | 60000 | - |
about, recombinant enzyme, gel filtration | Arabidopsis thaliana |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.4.99.5 | Chorismate | Arabidopsis thaliana | - |
Prephenate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.4.99.5 | Arabidopsis thaliana | P42738 | - |
- |
5.4.99.5 | Arabidopsis thaliana | Q9C544 | - |
- |
5.4.99.5 | Arabidopsis thaliana | Q9S7H4 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.4.99.5 | AtCM1, sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta II (DE3) | Arabidopsis thaliana |
5.4.99.5 | recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta II (DE3) by nickel affinity chromatography, tag cleavage with thrombin, dialysis, benzamidine/nickel affinity chromatography, and gel filtration | Arabidopsis thaliana |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.4.99.5 | Chorismate | - |
Arabidopsis thaliana | Prephenate | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
5.4.99.5 | homodimer | 2 * 32600, recombinant enzyme, SDS-PAGE | Arabidopsis thaliana |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.4.99.5 | AtCM1 | - |
Arabidopsis thaliana |
5.4.99.5 | AtCM2 | - |
Arabidopsis thaliana |
5.4.99.5 | AtCM3 | - |
Arabidopsis thaliana |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.4.99.5 | 13 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana | |
5.4.99.5 | 16.1 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana | |
5.4.99.5 | 38.7 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.4.99.5 | 8 | - |
assay at | Arabidopsis thaliana |
EC Number | General Information | Comment | Organism |
---|---|---|---|
5.4.99.5 | additional information | the crystal structure of AtCM1 in complex with tyrosine and phenylalanine identifies differences in the effector sites of the allosterically regulated yeast enzyme and the other two Arabidopsis isoforms. The catalytic efficiency (kcat/Km) of AtCM2 is 11 and 22fold higher than that of AtCM1 and AtCM3, respectively. This results from a combination of a more rapid turnover rate and a lower Km value for chorismate displayed by AtCM2 compared with the other two isoforms. Two catalytic residues (Arg229 and Lys240 in AtCM1) are invariant across the AtCM isoforms. These two basic residues are essential for substrate binding, orient the two negatively charged carboxylic acids of chorismate, and provide transition state stabilization during catalysis | Arabidopsis thaliana |
5.4.99.5 | physiological function | AtCM2 is a nonallosteric form | Arabidopsis thaliana |
5.4.99.5 | physiological function | isozyme AtCM1 is alosterically regulated | Arabidopsis thaliana |
5.4.99.5 | physiological function | isozyme AtCM3 is alosterically regulated | Arabidopsis thaliana |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.4.99.5 | 11.8 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana | |
5.4.99.5 | 29.27 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana | |
5.4.99.5 | 258 | - |
chorismate | recombinant enzyme, pH 8.0, temperature not specified in the publication | Arabidopsis thaliana |