Cloned (Comment) | Organism |
---|---|
overexpressed in Escherichia coli | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
using the hanging drop, vapor-diffusion method in the presence of 5 mM 3-phosphoshikimate | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
P101S | the substrate binding affinities, as reflected by the respective Km values, are only slightly decreased for the P101S mutant. Km (3-phosphoshikimate): 0.071 mM, Km (phosphoenolpyruvate): 0.071, kcat/Km (phosphoenolpyruvate): 230000/Msec, kcat/Km (3-phosphoshikimate): 240000/Msec. Mutant P101S is moderately inhibited by glyphosate | Escherichia coli |
T971I | the single site T97I mutation renders the enzyme sensitive to glyphosate and causes a substantial decrease in the affinity for phosphoenolpyruvate. Km (3-phosphoshikimate): 0.077 mM, Km (phosphoenolpyruvate): 0.38, kcat/Km (phosphoenolpyruvate): 23000/Msec, kcat/Km (3-phosphoshikimate): 1200000/Msec | Escherichia coli |
T97I/P101S | mutant is essentially insensitive to glyphosate (Ki 2.4 mM) but maintains high affinity for the substrate phosphoenolpyruvate (Km: 0.1 mM) and 3-phosphoshikimate (Km: 0.077 mM). kcat/Km (phosphoenolpyruvate): 57000/Msec, kcat/Km (3-phosphoshikimate): 740000/Msec. The crystal structure at 1.7 A resolution reveals that the dual mutation causes a shift of residue Gly96 toward the glyphosate binding site, impairing efficient binding of glyphosate, while the side chain of Ile97 points away from the substrate binding site, facilitating phosphoenolpyruvate utilization | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
glyphosate | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
phosphoenolpyruvate | wild-type | Escherichia coli | |
0.048 | - |
3-phosphoshikimate | wild-type | Escherichia coli | |
0.071 | - |
phosphoenolpyruvate | mutant P101S | Escherichia coli | |
0.071 | - |
3-phosphoshikimate | mutant P101S | Escherichia coli | |
0.077 | - |
3-phosphoshikimate | mutant P101S/T97I | Escherichia coli | |
0.077 | - |
3-phosphoshikimate | mutant T97I | Escherichia coli | |
0.1 | - |
phosphoenolpyruvate | mutant P101S/T97I | Escherichia coli | |
0.38 | - |
phosphoenolpyruvate | mutant T97I | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6D3 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
phosphoenolpyruvate + 3-phosphoshikimate | - |
Escherichia coli | phosphate + 5-enolpyruvylshikimate 3-phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
EPSPS | - |
Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0003 | - |
glyphosate | wild-type | Escherichia coli | |
0.003 | - |
glyphosate | mutant P101S | Escherichia coli | |
0.09 | - |
glyphosate | mutant P101S | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
23000000 | - |
phosphoenolpyruvate | mutant T97I | Escherichia coli | |
57000000 | - |
phosphoenolpyruvate | mutant P101S/T97I | Escherichia coli | |
74000000 | - |
3-phosphoshikimate | mutant P101S/T97I | Escherichia coli | |
120000000 | - |
3-phosphoshikimate | mutant T97I | Escherichia coli | |
240000000 | - |
3-phosphoshikimate | mutant P101S | Escherichia coli | |
250000000 | - |
phosphoenolpyruvate | mutant P101S | Escherichia coli | |
910000000 | - |
3-phosphoshikimate | wild-type | Escherichia coli | |
930000000 | - |
phosphoenolpyruvate | wild-type | Escherichia coli |