Information on EC 4.2.3.5 - chorismate synthase

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

EC NUMBER
COMMENTARY hide
4.2.3.5
-
RECOMMENDED NAME
GeneOntology No.
chorismate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
5-O-(1-carboxyvinyl)-3-phosphoshikimate = chorismate + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1,4-trans elimination
beta-elimination
-
-
elimination
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
chorismate biosynthesis from 3-dehydroquinate
-
-
chorismate metabolism
-
-
Metabolic pathways
-
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
5-O-(1-carboxyvinyl)-3-phosphoshikimate phosphate-lyase (chorismate-forming)
Requires FMN. The reaction goes via a radical mechanism that involves reduced FMN and its semiquinone (FMNH(.)). Shikimate is numbered so that the double-bond is between C-1 and C-2, but some earlier papers numbered the ring in the reverse direction.
CAS REGISTRY NUMBER
COMMENTARY hide
9077-07-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
Uniprot
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
pea
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
GenBank
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(6S)-6-fluoro-5-enolpyruvylshikimate 3-phosphate
6-fluorochorismate + phosphate
show the reaction diagram
-
converion rate between 270 and 370 times slower than natural substrate conversion
-
-
?
(6S)-6-fluoro-5-enolpyruvylshikimate-3-phosphate
6-fluorochorismate + phosphate
show the reaction diagram
-
?
-
-
5-enolpyruvylshikimate 3-phosphate
chorismate + phosphate
show the reaction diagram
5-enolpyruvylshikimate-3-phosphate
chorismate + phosphate
show the reaction diagram
-
-
-
-
?
5-O-(1-carboxyvinyl)-3-phosphoshikimate
chorismate + phosphate
show the reaction diagram
-
seventh enzyme of the shikimate pathway, catalyzes the NADH- and FMN-dependent synthesis of chorismate, a precursor of aromatic amino acids, naphthoquinones, menaquinones, and mycobactins
-
-
r
FMN + NADPH
?
show the reaction diagram
O5-(1-carboxyvinyl)-3-phosphoshikimate
?
show the reaction diagram
O5-(1-carboxyvinyl)-3-phosphoshikimate
chorismate + phosphate
show the reaction diagram
additional information
?
-
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
5-enolpyruvylshikimate 3-phosphate
chorismate + phosphate
show the reaction diagram
5-enolpyruvylshikimate-3-phosphate
chorismate + phosphate
show the reaction diagram
-
-
-
-
?
5-O-(1-carboxyvinyl)-3-phosphoshikimate
chorismate + phosphate
show the reaction diagram
-
seventh enzyme of the shikimate pathway, catalyzes the NADH- and FMN-dependent synthesis of chorismate, a precursor of aromatic amino acids, naphthoquinones, menaquinones, and mycobactins
-
-
r
O5-(1-carboxyvinyl)-3-phosphoshikimate
?
show the reaction diagram
O5-(1-carboxyvinyl)-3-phosphoshikimate
chorismate + phosphate
show the reaction diagram
additional information
?
-
-
the dissociation constant for the S127A mutant protein is similar to that of the wild-type enzyme, whereas the replacement of S16 with alanine results in a slight increase of the dissociation constant
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
flavins
additional information
-
no activity with 5-deaza-FMN
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(3R,4R,5R)-5-(carboxymethoxy)-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
-
(3R,4S,5R)-4-hydroxy-3-[(2S)-1-hydroxy-1-oxo-2-phosphonooxypropan-2-yl]oxy-5-phosphonooxycyclohexene-1-carboxylic acid
-
(3R,4S,5R)-5-(1-carboxyethoxy)-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
-
(3R,4S,5R)-5-[(1R)-1-carboxy-1-phosphonoethoxy]-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
-
(3R,4S,5S,6S)-5-(1-carboxyethenoxy)-6-fluoro-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
-
(6R)-6-fluoro-5-enoylpyruvylshikimate 3-phosphate
-
IC50 is 0.0005 mM when the enzyme is preincubated with the inhibitor, the IC50 is 0.25 mM when the enzyme is not preincubated with the inhibitor, inhibition is not absolutely irreversible
(6S)-6-fluoro-5-enolpyruvylshikimate 3-phosphate
-
competitive inhibitor
2-difluoro-[1-phosphonooxyethoxy]-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
-
bathophenanthroline
-
-
diethyl dicarbonate
-
-
dioxygen
-
inactivation
O2
-
activity only in anaerobic conditions
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
light
-
stimulation
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0027 - 0.08
5-enolpyruvylshikimate 3-phosphate
0.043 - 0.28
NADPH
0.0013 - 0.08
O5-(1-carboxyvinyl)-3-phosphoshikimate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.012 - 0.87
5-enolpyruvylshikimate 3-phosphate
0.0083
NADH
Mycobacterium tuberculosis
-
kinetics of reduction of the MtCS-bound FMNox is determined using fixed amount of holoenzyme and varying levels of NADH
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00179
(3R,4R,5R)-5-(carboxymethoxy)-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
theoretical Ki value obtained using AUTODOCK software version 4, pH not specified in the publication, temperature not specified in the publication
0.000109
(3R,4S,5R)-4-hydroxy-3-[(2S)-1-hydroxy-1-oxo-2-phosphonooxypropan-2-yl]oxy-5-phosphonooxycyclohexene-1-carboxylic acid
theoretical Ki value obtained using AUTODOCK software version 4, pH not specified in the publication, temperature not specified in the publication
0.000192
(3R,4S,5R)-5-(1-carboxyethoxy)-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
theoretical Ki value obtained using AUTODOCK software version 4, pH not specified in the publication, temperature not specified in the publication
0.000526
(3R,4S,5R)-5-[(1R)-1-carboxy-1-phosphonoethoxy]-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
theoretical Ki value obtained using AUTODOCK software version 4, pH not specified in the publication, temperature not specified in the publication
0.000815
(3R,4S,5S,6S)-5-(1-carboxyethenoxy)-6-fluoro-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
theoretical Ki value obtained using AUTODOCK software version 4, pH not specified in the publication, temperature not specified in the publication
0.00061
2-difluoro-[1-phosphonooxyethoxy]-4-hydroxy-3-phosphonooxycyclohexene-1-carboxylic acid
theoretical Ki value obtained using AUTODOCK software version 4, pH not specified in the publication, temperature not specified in the publication
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.25
(6R)-6-fluoro-5-enoylpyruvylshikimate 3-phosphate
Escherichia coli
-
IC50 is 0.0005 mM when the enzyme is preincubated with the inhibitor, the IC50 is 0.25 mM when the enzyme is not preincubated with the inhibitor, inhibition is not absolutely irreversible
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.004
-
CS activity
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
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enzyme form CS1; enzyme form CS2
6.5 - 8.5
-
-
7.5
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
parasite cytosol
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Bifidobacterium longum subsp. infantis (strain ATCC 15697 / DSM 20088 / JCM 1222 / NCTC 11817 / S12)
Campylobacter jejuni subsp. jejuni serotype O:2 (strain ATCC 700819 / NCTC 11168)
Helicobacter pylori (strain ATCC 700392 / 26695)
Helicobacter pylori (strain ATCC 700392 / 26695)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
41800
-
determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis and electrospray mass spectrometry
80100
-
gel filtration
110000 - 138000
-
gel filtration
110000
-
sucrose density gradient centrifugation
144000
-
gel filtration after cross-linkage with dimethyl suberimidate
186800
-
gel filtration
190000
-
nondenaturing PAGE
198000
200000
non-denaturing PAGE in presence of O5-(1-carboxyvinyl)-3-phosphoshikimate and FMN
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
trimer
heterotrimer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of the enzyme reveals a novel beta alpha beta sandwich topology
crystal structure of chorismate synthase in both FMN-bound and FMN-free form. It is a tetrameric enzyme, with each monomer possessing a novel beta-alpha-beta sandwich fold
hanging-drop vapour-diffusion method, crystallization at 296 K using polyethylene glycol 400 as precipitant, recombinant enzyme fused with an eight-residue C-terminal tag
-
beta-alpha-beta sandwich structure
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homology modeling of structure of chorismate synthase along with cofactor FMN using crystal structure of Helicobacter pylori chorismate synthase. Each monomer of Plasmodium falciparum chorismate synthase consists of 10 helices and 13 sheets. The core of monomer has a unique beta-alpha-beta sandwich fold that appears as a three layered structure in the predicted model. This unique fold consists of two antiparallel five stranded beta-sheet layers and four alpha-helices which are sandwiched between these two beta-sheet layers. The unique beta-alpha-beta-sandwich provides a scaffold for cofactor and substrate binding. The residues involved in polar interactions with substrate 5-enolpyruvylshikimate-3-phosphate are Arg46, Lys60, Asp61, Lys90, Ser121, Ser122 and Arg491
hanging drop vapour diffusion method, three-dimensional X-ray structure from selenomethionine-labeled crystals at 2.2 A resolution. The structure shows a novel beta,alpha,beta,alpha fold consisting of an alternate tight packing of two alpha-helical and two beta-sheet layers. The molecule is arranged as a tight tetramer with D2 symmetry
hanging-drop vapour diffusion method. Crystal streucture solved at 1.0 A
-
modelling the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate in chorismate synthase unsing B3LYP density functional theory and ab initio QM/MM methods, based on PDB entry 1QXO. In the reaction mechanism phosphate elimination precedes proton transfer. B3LYP predicts reaction energetics that are qualitatively wrong
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
92
melting temperature above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
apparent stability of all mutant proteins is comparable to that of the wild-type enzyme
-
bovine serum albumin stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, 0.1 M potassium phosphate buffer, pH 7.0, 0.1 mM EDTA, 0.2 mM dithiothreitol, more than 6 months
-
-15C, several months
-
-20C, 0.05 mM Tris-HCl, pH 7.5, 0.01 mM FMN, 10% glycerol, 3 mM mercaptoethanol, 4 months
-
-20C, 50 mM Tris-HCl buffer, pH 7.5, 0.4 mM dithiothreitol, 50% glycerol
4C, 2 weeks
-
liquid nitrogen
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by two-step chromatographic procedure
-
by using a 3-step purification (anion exchange, hydrophobic interaction and anion exchange chromatography)
-
expressed in Escherichia coli
recombinant enzyme
recombinant enzyme fused with an eight-residue C-terminal tag
-
together with NADPH dependent flavin reductase
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and expression in Escherichia coli strain Rosetta(DE3) in the absence of IPTG induction, heterologous overexpression of the MtCS protein
-
expressed in Saccharomyces cerevisiae
expression in a chorismate synthase deficient Saccharomyces cerevisiae strain
expression in Escherichia coli
mutant proteins are heterologously expressed in Escherichia coli, strain BL21(DE3)RP
-
overexpression in Escherichia coli
overexpression in Escherichia coli, fused with an eight-residue C-terminal tag
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D367A
-
comparable to the wild-type enzyme with respect to substrate and cofactor binding, but activity significantly lower
D367N
-
comparable to the wild-type enzyme with respect to substrate and cofactor binding, but activity significantly lower
S127A
-
amino acid replacements are performed using the QuikChange site-directed mutagenesis kit
S16A
-
amino acid replacements are performed using the QuikChange site-directed mutagenesis kit
S16A/S127A
-
double-mutant protein
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
-
enzymes of shikimate pathway are attractive targets to the development of antitubercular agents
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