Information on EC 2.5.1.19 - 3-phosphoshikimate 1-carboxyvinyltransferase and Organism(s) Escherichia coli and UniProt Accession P0A6D3

for references in articles please use BRENDA:EC2.5.1.19
Word Map on EC 2.5.1.19
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
This record set is specific for:
Escherichia coli
UNIPROT: P0A6D3
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota


The taxonomic range for the selected organisms is: Escherichia coli

EC NUMBER
COMMENTARY hide
2.5.1.19
-
RECOMMENDED NAME
GeneOntology No.
3-phosphoshikimate 1-carboxyvinyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
phosphoenolpyruvate + 3-phosphoshikimate = phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
enolpyruvate group transfer
-
induced-fit mechanism, formation of a 2-(S) configured tetrahedral reaction intermediate by covalent linkage of the two substrates
enolpyruvate group transfer
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
chorismate biosynthesis from 3-dehydroquinate
-
-
chorismate metabolism
-
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
-
Metabolic pathways
-
-
Biosynthesis of secondary metabolites
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphoenolpyruvate:3-phosphoshikimate 5-O-(1-carboxyvinyl)-transferase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9068-73-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
30 min, 25°C, pH 7.5, in presence of 2 mM dithiothreitol
detection of phosphate generation with Lanzetta reagent and change in absorbance at 650 nm
-
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
-
via formation of a 2-(S)-tetrahedral reaction intermediate (TI), pH 7.5, 25°C, 30 min
examination by malachite green phosphate-release assay
-
?
5-enolpyruvylshikimate 3-phosphate + H2O
5-enolpyruvylshikimate 3-phosphate ketal
show the reaction diagram
-
enzymatic hydrolysis, pH 7.0, 25°C, in presence of phosphate scavenging system and excess of enzyme
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
?
show the reaction diagram
-
-
-
-
?
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
-
-
detection of inorganic phosphate by reaction with Lanzetta reagent and change in optical density at 660 nm
-
?
shikimate 3-phosphate + phosphoenolpyruvate
phosphate + 5-enpolpyruvylshikimate 3-phosphate
show the reaction diagram
-
enolpyruvylshikimate 3-phosphate ketal formation from shikimate 3-phosphate (radiolabelled) and phosphoenolypyruvate (in excess over shikimate 3-phosphate) by hydrolysis of product 5-enpolpyruvylshikimate 3-phosphate in presence of 25% excess of enzyme, pH 7.0, 25°C
analysis of products by ion-paired C18 reverse-phase HPLC and scintillation counting
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-enolpyruvylshikimate 3-phosphate
show the reaction diagram
phosphoenolpyruvate + 3-phosphoshikimate
phosphate + 5-O-(1-carboxyvinyl)-3-phosphoshikimate
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(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 enzyme’s backbone and active site (e.g., Glu341 and Arg124)
(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
(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
(3R,4S,5R)-5-((R)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
analogue of the tetrahedral reaction intermediate, competitive to both substrates, binding structure analysis, binding induces conformational changes to residues Arg124 and Glu341 within the active site, which results in structural alterations in the amino-terminal globular domain of the enzyme
(3R,4S,5R)-5-((S)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
analogue of the tetrahedral reaction intermediate, competitive to both substrates, binding structure analysis, binding induces no conformational changes
(Z)-3-fluorophosphoenolpyruvate
-
competitive vs. phosphoenolpyruvate at saturated shikimate 3-phosphate concentration and vice versa
3-Bromopyruvate
-
0.1 mM, approx. 80% inactivation after 5 min, maximum rate-constant: 0.31/min, substrates or a combination of shikimate 3-phosphat and glyphosate protect from inactivation, bromopyruvate modifies residues C408 and L411
5-Deoxy-shikimate 3-phosphate
-
competitive vs. shikimate 3-phosphate
5-enolpyruvylshikimate 3-phosphate
-
product inhibition
Carboxyallenyl phosphate
-
strong
diethyldicarbonate
-
inactivation with a second-order rate constant of 220/M/min, subtstrates protect from inactivation, enzyme activity is recovered by treatment with hydroxylamine
glyphosate
N-phosphonomethylglycine
phosphoenolpyruvate
-
substrate inhibition
pyruvate
-
20 mM, 85% inactivation after 1 h in the presence of cyanoborhydride, no inactivation in the absence of cyanoborhydride, preincubation with 5-enolpyruvylshikimate or a combination of shikimate 3-phosphate and glyphosate prevents from inactivation
additional information
-
insensitive to inhibition by glyphosate.
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.06 - 0.15
phosphoenolpyruvate
0.06 - 0.1
shikimate 3-phosphate
0.0025 - 2
3-phosphoshikimate
0.003 - 0.011
5-enolpyruvylshikimate 3-phosphate
1.2 - 4.6
phosphate
0.0024 - 22.5
phosphoenolpyruvate
0.02 - 0.135
shikimate 3-phosphate
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.026 - 100
3-phosphoshikimate
0.000012
5-enolpyruvylshikimate 3-phosphate
-
5-enpolpyruvylshikimate 3-phosphate ketal formation by hydrolysis of radiolabelled 5-enpolpyruvylshikimate 3-phosphate, pH 7.0, 25°C
0.0000045 - 100
phosphoenolpyruvate
additional information
phosphoenolpyruvate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0016 - 910000000
3-phosphoshikimate
0.0007 - 930000000
phosphoenolpyruvate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
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
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
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
0.0004 - 0.066
glyphosate
0.000016
(3R,4S,5R)-5-((R)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
versus 3-phosphoshikimate
0.00075
(3R,4S,5R)-5-((S)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid
-
versus 3-phosphoshikimate
0.0029 - 0.0064
(Z)-3-fluorophosphoenolpyruvate
0.0126
5-Deoxy-shikimate 3-phosphate
-
-
0.0003 - 0.09
glyphosate
0.0008 - 0.96
N-phosphonomethylglycine
additional information
glyphosate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05 - 8.2
glyphosate
additional information
glyphosate
Escherichia coli;
-
more than 10 mM for mutant G96A; more than 10 mM for mutant G96A/A183T
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8
-
+/-0.2 micromol/min/mg, mutant P101L
22
-
+/-0.4 micromol/min/mg, mutant P101S
28
-
+/-1 micromol/min/mg, mutant P101G
31
-
+/-0.5 micromol/min/mg, mutant P101A
50
-
+/-1 micromol/min/mg
0.0064
-
recombinant enzyme, activity in crude extracts
0.0076
-
wild-type enzyme, cell exract
0.0357
-
mutant T42M, cell extract
18.9
-
mutant A183T, with respect to phosphoenolpyruvate
20
-
recombinant His-tagged wild-type enzyme
21.1
-
phosphate + 5-enolpyruvylshikimate 3-phosphate, recombinant enzyme
24
-
recombinant wild-type enzyme
26.5
-
mutant G96A/A183T, with respect to phosphoenolpyruvate
34
-
with respect to phosphoenolpyruvate
37.4
-
mutant G96A, with respect to phosphoenolpyruvate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at
7.8
-
wild-type enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.6 - 8.4
-
H385N mutantt enzyme
6 - 8.4
-
wild-type enzyme
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
42000
-
gel filtration
44000
-
SDS-PAGE
45000
-
* 45000, wild-type and mutant enzyme, SDS-PAGE
46110
-
calculated from polypeptide sequence of aroA gene product
46112
-
1 * 46112, deduced from nucleotide sequence
49000
-
1 * 49000
55000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
* 45000, wild-type and mutant enzyme, SDS-PAGE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
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
-
mutants P101S and P101L in complex with shikimate 3-phosphate and with or without inhibitor glyphosate, long-range structural change in glyphosate binding site compared to wild-type, Gly96 and Thr97 are shifted towards the glyphosate-binding site, which gets slightly narrowed, crystals: space group: P2(1)2(1)2(1), unit cell parameters: a: 57.6-57.8, b: 85.1-85.6, c: 87.9-88.3, beta: 90°, hanging-drop vapour-diffusion, protein solution (37.5 mg/ml, 5 mM inhibitor and/or 5 mM shikimate 3-phosphate), 19°C, molecular replacement using PDB: 1G6S as model
-
enzyme complexed with inhibitor (3R,4S,5R)-5-((S)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid or with inhibitor (3R,4S,5R)-5-((R)-1-carboxy-1-phosphono-ethoxy)-4-hydroxy-3-phosphonooxy-cyclohex-1-enecarboxylic acid, 100 mg/ml recombinant enzyme in 2.5 M sodium formate with 10 mM inhibitor, hanging drop vapour diffusion method, 19°C, X-ray diffraction at -180°C using the rotation method on single flash-frozen crystals, structure determination and analysis at 1.5 and 1.9 A resolution, respectively
-
enzyme in complex with shikimate/glyphosate at 1.5 A resolution
-
purified recombinant wild-type and mutant enzymes in complex with the substrates, at 19°C, from 4 M sodium formate in presence of 5 mM 3-phosphoshikimate and 5 mM phosphoenolpyruvate, X-ray diffraction structure determination and analysis at 1.6 A resolution
semiempirical molecular modelling using, among others, the crystal structure of EPSPS mutant D313A (PDB: 1Q36) as model for assignment of protonation states of all basic amino acids in the active site: in the enzyme-tetrahedral reaction intermediate (TI) complex is residue His385 in a neutral form (with protonated epsilon-N atom) while residues Lys22, Lys340 and Lys411 are protonated, hydrogen bonds occur between Lys22 and the carboxylate oxygen atom of the phosphoenolpyruvate moiety of the TI, Asp313 has only minor effects on TI positioning within the active site but mediates as a base the attack of the TI C4-hydroxyl group on the TI methyl group prior to EPSP formation
-
using the hanging drop, vapor-diffusion method in the presence of 5 mM 3-phosphoshikimate
-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
complete unfolding of the protein at 2.5 M guanidine hydrochloride at pH 7.8 (midpoint guanidine hydrochloride concentration of unfolding: 0.8 +/-0.1 M), enhanced flexibility, lower stability and increased proteolysis of mutants A183T and G96A/A183T compared to wild-type and mutant G96A as revealed by susceptibility to proteolytic digestion with trypsin, thermolysin, pronase (25°C for various time intervals), unfolding profiles (intrinsic tryptophan fluorescence measurements at 340 nm) after treatment with guanidine hydrochloride (30 min at 25°C) and dynamic quenching with acrylamide or not, and secondary structure determination by circular dichroism spectroscopy
-
dithiothreitol reverses aggregation which sometimes occurs after storage at -20°C
-
dithiothreitol, 0.4 mM, stabilizes during purification
-
glycerol, 50% v/v, stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50 mM Tris-HCl, pH 7.5, 50 mM KCl, 0.4 mM dithiothreitol, 50% v/v glycerol, at least 4 months, no loss of activity
-
-20°C, in 50% v/v glycerol, enzyme concentration 5 mg/ml, at least 1 year
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from supernatant from 25% (w/v) ammonium sulfate precipitation of crude bacterial extracts, Q-sepharose anion exchange chromatography (NaCl gradient elution) and FPLC (Mono-Q column, NaCl gradient elution)
-
recombinant enzyme
-
recombinant enzyme, ammonium sulfate, DEAE-Sephacel, phenyl-Sepharose, phosphocellulose chromatography
-
recombinant wild-type and G96A mutant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
wild-type and mutants in pET-24d for expression in Escherichia coli BL21(DE3)
-
Escherichia coli structural gene aroA
-
Escherichia coli structural gene aroA; subcloned from bacteriophage lambdapserC into multicopy plasmid pAT153 with subsequent transformation of Escherichia coli AB2829 CGSC2829 cells
-
expressed as a His-tagged fusion protein
-
expression of the wild-type enzyme and of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium in strain XL1-Blue
-
expression of the wild-type enzyme, of the chimeric mutant aroA-M1, comprising N-terminal part of the Escherichia coli enzyme and the C-terminal part of the enzyme from Salmonella typhimurium, and of T42M-aroA mutant in strain BL21(DE3)
-
expression of wild-type and chimeras of Salmonella and Escherichia coli mutant enzymes in Escherichia coli
-
expression of wild-type and G96A mutant enzyme in Escherichia coli
-
in pET24d for expression in Escherichia coli DE3
-
mutant enzyme G96A/A183T is transferred to Brassica napus via Agrobacterium-mediated transformation. Transgenic plants are resistant to glyphosate at a concentration of 10 mM whereas the non-transformed control plants are unable to survive 1 mM glyphosate
-
overexpressed in Escherichia coli
-
overexpression in Escherichia coli
-
overexpression of wild-type and mutant enzymes in strain BL21(DE3)
overexpression of wild-type, His-tagged wild-type and several mutant enzymes in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
P101A
-
slight decrease in catalytic efficiency, decreased inhibitory potency of glyphosate
P101G
-
slight decrease in catalytic efficiency, decreased inhibitory potency of glyphosate
P101L
-
lowest catalytic efficiency, decreased inhibitory potency of glyphosate due to long-range conformational changes
P101S
-
slight decrease in catalytic efficiency, decreased inhibitory potency of glyphosate due to long-range conformational changes, flexibility of Ser101 side chain disables hydrophobic bonding with carbonyl oxygen of Thr97
A183T
-
38fold decrease in phosphoenolpyruvate-binding affinity, more solvent-exposed tryptophan residues and lower stability against guanidine hydrochloride compared to wild-type and mutant G96A, midpoint guanidine hydrochloride concentration of unfolding: 0.7 M, higher structural flexibility and decrease of secondary structure (28% alpha helix, 35% beta sheet) compared to wild-type (40% alpha helix, 31% beta sheet) and lowest resistance against proteolysis, residue A183 located on the exterior in the N-terminal domain
D242A
-
site-directed mutagenesis, the mutation is responsible for the high increase in activity
D313C
-
compared to wild-type mutations of D313 causes kcat to decrease. In the mutant D313C the kcat is smaller than in other mutants (1200fold). Cys is ionizable and can potentially act as an acid/base catalyst, or the thiolate form can stabilize cationic intermediates electrostatically. This accounts for the higher catalytic activity for D313C than for D313A: The effects on Km(3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest
D313L
-
compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold
D348A
-
site-directed mutagenesis, the mutation is responsible for the high increase in activity
D49A
-
41% of wild-type activity
E341C
-
compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold
E341M
-
compared to wild-type mutations of D313 causes kcat to decrease up to 30000fold while the effects on Km (3-phosphoshikimate) or Km (phosphoenolpyruvate) are modest, never more than 40fold
G96A/A183T
K340A
-
2.4% of wild-type activity
K411A
-
10.4% of wild-type activity
N94A
-
50% of wild-type activity
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
Q171A
-
1.7% of wild-type activity
R100A
-
site-directed mutagenesis, the mutation is responsible for the high increase in activity
R100M
-
0.2% of wild-type activity
R124A
-
19.6% of wild-type activity
R27A
-
binding of shikimate 3-phosphate is abolished
R344K
-
31.7% of wild-type activity
R344M
-
16.3% of wild-type activity
R386M
-
15.8% of wild-type activity
T42M
-
site-directed mutagenesis, the mutation is responsible for the glyphosate resistance and the low Km for phosphoenolpyruvate
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
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
Y200F
-
1% of wild-type activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
drug development
-
protonated enolpyruvylshikimate 3-phosphate (cation) intermediate as potential target for inhibitor design
synthesis
-
enzyme is a target for development and synthesis of antimicrobial drugs