Application | Comment | Organism |
---|---|---|
drug development | differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation | Staphylococcus aureus |
drug development | differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation | Escherichia coli |
drug development | differential inhibition of class I and class II EPSPS by tetrahedral reaction intermediate-analogues possibly due to alteration of open-close transition during catalysis and/or upon inhibitor binding but not due to energy differences during complex formation | Agrobacterium sp. |
Crystallization (Comment) | Organism |
---|---|
in presence of 10 mM inhibitor (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (2F-TI, PDB: 2PQ9), 2F-TI-binding induces large conformational change of Glu341 side chain compared to TI-binding, crystals: space group P2(1)2(1)2(1), unit cell parameters: a: 58.2, b: 85.1, c: 87.6, beta: 90°, hanging-drop vapour-diffusion, sodium format conditions, molecular replacement using PDB: 1G6S as model | Escherichia coli |
wild-type in presence of 5 mM inhibitor (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (2F-TI, PDB: 2PQB) or (3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (RP-TI, PDB: 2PQC), and mutant A100G in presence of 5 mM inhibitor 2F-TI (PDB: 2PQD): 2F-TI-binding induces only slight shift of Glu354 side chain but substantial conformational change of 2F-TI molecule that is independent of residue 100 but mediated by residue Thr101 and due to rigidity of active site, RP-TI-binding induces slight movement of Glu354 side chain and occurs via salt bridge with residue Arg128 but does not alter enzymes backbone compared to the shikimate 3-phosphate bound enzyme, crystals: space group P2(1), unit cell parameters: a: 62.8-63.4, b: 44.9-45.0, c: 77.1-77.6, beta: 106.1-106.3°, hanging-drop vapour-diffusion, ammonium sulfate conditions, molecular replacement using PDB: 2GG6 as model, co-crystallization with (3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid (SP-TI) not succeeded, docking of SP-TI into substrate- or inhibitor-bound structures indicates low electrostatic interactions between enzyme and inhibitor | Agrobacterium sp. |
Protein Variants | Comment | Organism |
---|---|---|
A100G | glyphosate-sensitive, compared to wild-type similar inhibition constants for (3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid, (3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid, and (3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | Agrobacterium sp. |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | RP-TI, (R)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 460fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli | Agrobacterium sp. | |
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | RP-TI, (R)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, very potent competitive inhibitor of EPSPS forward reaction, binding induces substantial conformational change in enzymes backbone and active site (e.g., Glu341 and Arg124) | Escherichia coli | |
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | RP-TI, (R)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 120-fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli | Staphylococcus aureus | |
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | SP-TI, (S)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, non-potent competitive inhibitor of EPSPS forward reaction | Agrobacterium sp. | |
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | SP-TI, (S)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, moderate competitive inhibitor of EPSPS forward reaction | Escherichia coli | |
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | SP-TI, (S)-phosphonate analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction | Staphylococcus aureus | |
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | 2F-TI, (R)-difluoromethyl analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 8fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli | Agrobacterium sp. | |
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | 2F-TI, (R)-difluoromethyl analogue of the (S)-tetrahedral reaction intermediate, very potent competitive inhibitor of EPSPS forward reaction | Escherichia coli | |
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | 2F-TI, (R)-difluoromethyl analogue of the (S)-tetrahedral reaction intermediate, competitive inhibitor of EPSPS forward reaction, 4-fold decrease in potency compared to glyphosate-sensitive class I EPSPS from Escherichia coli | Staphylococcus aureus | |
glyphosate | - |
Agrobacterium sp. | |
glyphosate | - |
Escherichia coli | |
glyphosate | - |
Staphylococcus aureus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Agrobacterium sp. | Q9R4E4 | - |
- |
Agrobacterium sp. CP4 | Q9R4E4 | - |
- |
Escherichia coli | P0A6D3 | - |
- |
Staphylococcus aureus | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
shikimate 3-phosphate + phosphoenolpyruvate | via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25°C, 20 min | Staphylococcus aureus | phosphate + 5-enpolpyruvylshikimate 3-phosphate | examination by malachite green phosphate-release assay | ? | |
shikimate 3-phosphate + phosphoenolpyruvate | via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25°C, 30 min | Escherichia coli | phosphate + 5-enpolpyruvylshikimate 3-phosphate | examination by malachite green phosphate-release assay | ? | |
shikimate 3-phosphate + phosphoenolpyruvate | via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25°C, 30 min | Agrobacterium sp. | phosphate + 5-enpolpyruvylshikimate 3-phosphate | examination by malachite green phosphate-release assay | ? | |
shikimate 3-phosphate + phosphoenolpyruvate | via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25°C, 30 min | Agrobacterium sp. CP4 | phosphate + 5-enpolpyruvylshikimate 3-phosphate | examination by malachite green phosphate-release assay | ? |
Synonyms | Comment | Organism |
---|---|---|
5-enolpyruvylshikimate 3-phosphate synthase | class I, glyphosate-sensitive | Escherichia coli |
5-enolpyruvylshikimate 3-phosphate synthase | class II, glyphosate-tolerant | Staphylococcus aureus |
5-enolpyruvylshikimate 3-phosphate synthase | class II, glyphosate-tolerant | Agrobacterium sp. |
EPSP synthase | - |
Staphylococcus aureus |
EPSP synthase | - |
Escherichia coli |
EPSP synthase | - |
Agrobacterium sp. |
EPSPS | - |
Staphylococcus aureus |
EPSPS | - |
Escherichia coli |
EPSPS | - |
Agrobacterium sp. |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000039 | - |
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-0.6 nM | Escherichia coli | |
0.0000078 | - |
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-0.5 nM | Escherichia coli | |
0.00003 | - |
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-6 nM | Staphylococcus aureus | |
0.000063 | - |
(3R,4S,5R)-5-[(2R)-1,1-difluoro-3-hydroxy-3-oxo-2-phosphonooxy-propan-2-yl]oxy-4-hydroxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-15 nM | Agrobacterium sp. | |
0.00045 | - |
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-70 nM | Staphylococcus aureus | |
0.00076 | - |
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-200 nM | Escherichia coli | |
0.0018 | - |
(3R,4S,5R)-4-hydroxy-5-[(2R)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-0.0006 mM | Agrobacterium sp. | |
0.011 | - |
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-0.002 mM | Staphylococcus aureus | |
0.076 | - |
(3R,4S,5R)-4-hydroxy-5-[(2S)-1-hydroxy-1-oxo-2-phosphono-propan-2-yl]oxy-3-phosphonooxy-cyclohexene-1-carboxylic acid | +/-0.009 mM | Agrobacterium sp. |