Information on EC 5.4.99.5 - Chorismate mutase

New: Word Map on EC 5.4.99.5
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
5.4.99.5
-
RECOMMENDED NAME
GeneOntology No.
Chorismate mutase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Chorismate = prephenate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
group transfer
-
-
intramolecular
-
isomerization
rearrangement
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
bacilysin biosynthesis
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
L-phenylalanine biosynthesis I
-
-
L-phenylalanine biosynthesis II
-
-
L-tyrosine biosynthesis I
-
-
L-tyrosine biosynthesis II
-
-
L-tyrosine biosynthesis III
-
-
Metabolic pathways
-
-
phenylalanine metabolism
-
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
-
salinosporamide A biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
Chorismate pyruvatemutase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9068-30-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
bifunctional enzyme: chorismate mutase/prephenate dehydratase
-
-
Manually annotated by BRENDA team
Anthophyta
contain 3 isoenzymes with the exception of some closely related Leguminosae
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain 23 and its derivatives have 3 distinct enzyme species: CM1, CM2, and CM3. Strain 168 has only the CM3 form. Enzyme forms CM1 and CM2 may represent different aggregation states involving at least one common subunit
-
-
Manually annotated by BRENDA team
bifunctional enzyme: chorismate mutase/prephenate dehydratase
-
-
Manually annotated by BRENDA team
Pb 156, 2 enzyme forms: MW 55000 and MW 59000
-
-
Manually annotated by BRENDA team
Claviceps sp.
SD 58
-
-
Manually annotated by BRENDA team
Claviceps sp. SD 58
SD 58
-
-
Manually annotated by BRENDA team
basonym Alcaligenes eutrophus
-
-
Manually annotated by BRENDA team
Orchard grass
-
-
Manually annotated by BRENDA team
strain JFM-30
-
-
Manually annotated by BRENDA team
strain JM101
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
enzyme forms: CM1, CM2, and CM3
-
-
Manually annotated by BRENDA team
nonpathogenic species, Analysis of promoter activity described
Swissprot
Manually annotated by BRENDA team
Nephrolysis sp.
2 isoenzymes: CM1, CM2
-
-
Manually annotated by BRENDA team
one enzyme form: CM1
-
-
Manually annotated by BRENDA team
2 enzyme forms: CM1 and CM2
-
-
Manually annotated by BRENDA team
no activity in Corynebacterium diphtheriae
-
-
-
Manually annotated by BRENDA team
no activity in Mycobacterium leprae
-
-
-
Manually annotated by BRENDA team
no activity in Nocardia farcinica
-
-
-
Manually annotated by BRENDA team
-
Uniprot
Manually annotated by BRENDA team
2 enzyme forms: CM1 and CM3
-
-
Manually annotated by BRENDA team
Penicillium duponti
3 enzyme forms: CM1, CM2, CM3
-
-
Manually annotated by BRENDA team
cultivar Mitchell Diploid
UniProt
Manually annotated by BRENDA team
Pinus sp.
2 isoenzymes: CM1, CM2
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
2 isoenzymes: CM1 and CM2
-
-
Manually annotated by BRENDA team
Selaginella sp.
2 isoenzymes: CM1, CM2
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Streptomyces aureofaciens tue 24
tue 24
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Xanthomonas oryzae pv. oryzae XKK.12
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
additional information
-
structure-function relationships of chorismate-utilizing enzymes, structure comparisons, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
carbachorismate
carbaprephenate
show the reaction diagram
-
-
-
?
Chorismate
?
show the reaction diagram
Chorismate
Prephenate
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
Chorismate
?
show the reaction diagram
Chorismate
Prephenate
show the reaction diagram
additional information
?
-
-
isochorismate-pyruvate lyase, PchB EC 4.2.99.21, can also perform the chorismate mutase reaction
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
-
enhances activity
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(1R,2S,3S,5S,7S)-10-hydroxy-3-oxo-2-oxa-5-azatricyclo[4.3.1.1(4,8)]undecane-8-carboxylate, sodium salt
-
does not display tighter binding to the enzyme than the native substrate chorismate or greater inhibitory action than the ether analogue
-
(1R,3R,5S)-3-carboxy-1-hydroxy-2-oxabicyclo[3.3.1]non-6-ene-5-carboxylate
(1R,3R,5S,8R)-2-azatricyclo[3.3.1.0(1,8)]-non-6-ene-3,5-dicarboxylate, disodium salt
-
exo arizidine analogue, no time-dependent loss of activity is observed in the presence of this potentially reactive aza inhibitor
-
(1R,3R,5S,8S)-8-hydroxy-2-azabicyclo[3.3.1]non-6-ene-3,5-dicarboxylate, disodium salt
-
does not display tighter binding to the enzyme than the native substrate chorismate or greater inhibitory action than the ether analogue
-
(1R,3S,5S,8S)-8-hydroxy-2-azabicyclo[3.3.1]non-6-ene-3,5-dicarboxylate, disodium salt
-
does not display tighter binding to the enzyme than the native substrate chorismate or greater inhibitory action than the ether analogue
-
(1S,3R,5R)-1-hydroxy-5-nitro-2-oxabicyclo[3.3.1]non-6-ene-3-carboxylic acid
(1S,3S,5R)-1-hydroxy-5-nitro-2-oxabicyclo[3.3.1]non-6-ene-3-carboxylic acid
(1S,3S,5R,6R)-6-hydroxy-4-oxabicyclo[3.3.1]non-7-ene-1,3-dicarboxylate
-
endo-oxabicyclic dicarboxylic acid is a good geometric mimic of transition state
(1S,4S,6R,8S,10S)-3-oxo-5-aza-2-oxa-tetracyclo[4.3.1.(4,8).0(6,10)]undecane-8-carboxylate, sodium salt
-
tetracyclic lactone, no time-dependent loss of activity is observed in the presence of this potentially reactive aza inhibitor
-
(2E)-8-exo-3-Hydroximino-8-hydroxy-2-oxabicyclo-[3.3.1]non-6-ene-5-carboxylic acid
-
poor
(2Z)-2-(4-chlorophenyl)-3-(4,5-dimethoxy-2-nitrophenyl)prop-2-enoic acid
-
-
(3R,6Z)-8-hydroxy-2-azabicyclo[3.3.3]undec-6-ene-3,5-dicarboxylic acid
-
-
(3S,6Z)-8-hydroxy-2-azabicyclo[3.3.3]undec-6-ene-3,5-dicarboxylic acid
-
-
(3S,6Z)-8-hydroxy-2-oxabicyclo[3.3.3]undec-6-ene-3,5-dicarboxylic acid
-
-
1-Substituted adamantane derivatives
-
order of decreasing inhibitory activity with the various substituents: -PO32-, -P(OCH3)O2, CO2-, -CH2CO2-, -SO2-,Y -SO3-
2-(1-Carboxy-1,4-dihydrobenzyl)acrylic acid
-
-
3-Chloroadamantane-1-acetic acid
-
-
3-endo,6-exo-6-Hydroxy-7-bicyclo[3.3.1]-nonene-1,3-dicarboxylic acid
-
poor
3-endo,8-exo-8-Hydroxy-2-oxabicyclo[3.3.1]non-6-ene-3,5-dicarboxylic acid
-
potent
4-Methyl-DL-Trp
-
enzyme form CM1 is inhibited, enzyme form CM2 not
4-[[2-(3,4-dimethoxyphenyl)ethyl]amino]-3-nitro-5-sulfamoylbenzoic acid
-
-
5,5'-dithiobis(2-nitrobenzoate)
-
-
6-Methyl-DL-Trp
-
enzyme form CM1 is inhibited, enzyme form CM2 not
8-exo-8-Hydroxy-2-oxabicyclo[3.3.1]nona-3,6-diene-3,5-dicarboxylic acid
-
slight
8-hydroxy-2-oxa-bicyclo[3.3.1]non-6-ene-3,5-dicarboxylic acid
-
competitive inhibition
-
Adamantane-1-acetic acid
-
-
adamantane-1-phosphonate
-
no inhibitory effect up to concentrations of 0.1 and 1 mM
caffeic acid
chlorogenic acid
-
enzyme forms CM1 and CM2 are inhibited, enzyme form CM3 is unaffected
chorismate
DL-3-fluoro-Phe
-
enzyme form CM1 is inhibited, enzyme form CM2 not
DL-5-Fluoro-Trp
-
enzyme form CM1 is inhibited, enzyme form CM2 not
DL-5-hydroxy-Trp
-
enzyme form CM1 is inhibited, enzyme form CM2 not
endo-Oxabicylic transition state analogue inhibitor
-
-
-
ferulic acid
-
inhibits enzyme form CM3
iodoacetamide
-
-
L-Trp
-
96% residual activity at 0.2 mM
L-tryptophan
methyl 4-(methylamino)-3-nitrobenzoate
-
-
N-[[3-(tert-butoxycarbonyl)-2-phenyl-1,3-thiazolidin-4-yl]carbonyl]leucine
-
-
NaCl
-
inhibition is cooperative, NaCl also increases the sensitivity of the enzyme to inhibition by Phe
oxabicyclic dicarboxylic acid
-
transition state analogon, competitive inhibition
p-coumaric acid
-
inhibits enzyme form CM1, CM2 and CM3
prephenate
Transition state analogue inhibitor
-
-
-
tyrosine
Zn2+
-
strong
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3,4-dimethoxycinnamic acid
3-deoxy-D-arabino-heptulosonate-7-phosphate synthase
arabinose
expression can be induced by arabinose under the control of the araBAD promoter
caffeic acid
tryptophan
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03 - 16
chorismate
0.47 - 1
prephenate
additional information
Chorismic acid
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0003 - 366000
chorismate
94100 - 94140
prephenate
additional information
2-[5-Amino-2-(4-fluoro-phenyl)-6-oxo-6H-pyrimidin-1-yl]-N-(1-benzyl-2-oxo-2-thiazol-2-yl-ethyl)-acetamide
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.19 - 1.75
chorismate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 0.051
(1R,3R,5S)-3-carboxy-1-hydroxy-2-oxabicyclo[3.3.1]non-6-ene-5-carboxylate
0.00032 - 0.0076
(1S,3R,5R)-1-hydroxy-5-nitro-2-oxabicyclo[3.3.1]non-6-ene-3-carboxylic acid
0.23
(1S,3S,5R)-1-hydroxy-5-nitro-2-oxabicyclo[3.3.1]non-6-ene-3-carboxylic acid
-
30C, pH 7.5, wild-type
0.01771
(2Z)-2-(4-chlorophenyl)-3-(4,5-dimethoxy-2-nitrophenyl)prop-2-enoic acid
-
-
0.0057
4-[[2-(3,4-dimethoxyphenyl)ethyl]amino]-3-nitro-5-sulfamoylbenzoic acid
-
4-[[2-(3,4-dimethoxyphenyl)ethyl]amino]-3-nitro-5-sulfamoylbenzoic acid shows the most potent inhibition with Ki value of 0.0057 mM against MtCM
0.0037
8-hydroxy-2-oxa-bicyclo[3.3.1]non-6-ene-3,5-dicarboxylic acid
-
-
-
0.0028
chorismate
30C, pH 7.6, in presence of 0.005 mM tyrosine
0.0288
methyl 4-(methylamino)-3-nitrobenzoate
-
-
0.0211
N-[[3-(tert-butoxycarbonyl)-2-phenyl-1,3-thiazolidin-4-yl]carbonyl]leucine
-
-
0.003 - 1.1
oxabicyclic dicarboxylic acid
0.07 - 0.17
prephenate
0.034
tyrosine
70C, pH 7.6
additional information
oxabicyclic dicarboxylic acid
-
>> 1 mM mutant C88S/R90K
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.007 - 18
Saccharomyces cerevisiae chorismate mutase inhibitors
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.44
-
cell-free supernatant
25
after affinity chromatography
48.6
-
after 110.4fold purification
168
Superdex 200pg pool
210
-
recombinant enzyme
1200
-
0.1 mM tyrosine, wild-type
3300
-
0.1 mM tyrosine, mutant T226D
3700
-
unliganded, mutant T226D
4800
-
unliganded, wild-type
8400
0.1 mM tyrosine, wild-type
11100
-
0.5 mM tryptophan, mutant T226D
13400
0.1 mM tyrosine, mutant D233I
20600
-
unliganded, mutant T226I
21700
-
0.1 mM tyrosine, mutant T226I
22000
0.1 mM tyrosine, mutant D233T
26400
-
0.5 mM tryptophan, mutant T226I
30300
unliganded, mutant D233I
32500
unliganded, wild-type
40200
-
0.5 mM tryptophan, wild-type
41300
unliganded, mutant D233T
63000
0.5 mM tryptophan, mutant D233I
64300
0.5 mM tryptophan, mutant D233T
88500
0.5 mM tryptophan, wild-type
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.4
catalytic maximum in presence of tyrosine
6 - 8
-
enzyme form CM1 and CM2
6 - 10
6.2
-
55000 MW enzyme form
6.4
-
low-activity strain 168
6.6 - 7
7.1
catalytic maximum in presence of tryptophan
7.5 - 9.5
-
enzyme form CM2
7.8
-
enzyme form CM1
8
-
in absence of Trp
8.9
-
high activity strain WB672
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35 - 37
-
enzyme form CM1
38
-
decrease in turnover at temperatures higher than 38C
48
maximum enzymatic activity
56
-
enzyme form CM2
70
-
As the temperature is lowered, distances of electrostatic and hydrophobic interactions decrease. The distance between Glu77-COOH and the hydroxyl oxygen decreases from the optimum
additional information
-
The average structures of the thermophilic E.S complex at three different temperatures are obtained from the MD simulations. A snapshot of the most important features of the active site of the TtCM E.S complex at 70C is shown. At this temperature the Boltzmann distribution is primarily composed of reactive conformers - near attack conformers (NACs). The effects of the different temperatures on the distances in the structure are shown. The distances between electrostatic and hydrophobic pairs decrease as temperature decreases
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 70
-
studies that elucidate the dependence of the E-S active site on temperatures 25, 60 and 70C
37 - 50
-
maximally active in this range
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.7 - 4.8
isoelectric focusing, pH-gradient 3.5-9.5
7.7
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
esophageal gland cell
Manually annotated by BRENDA team
-
all three enzyme forms are present: CM1, CM2, and CM3
Manually annotated by BRENDA team
subventral and dorsal pharyngeal glands; subventral and dorsal pharyngeal glands
Manually annotated by BRENDA team
-
enzyme forms are present: CM1, CM2, and CM3
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
isoform CM1 is localized to the chloroplast stroma
Manually annotated by BRENDA team
-
from mesophyll
-
Manually annotated by BRENDA team
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bartonella henselae (strain ATCC 49882 / DSM 28221 / Houston 1)
Burkholderia thailandensis (strain E264 / ATCC 700388 / DSM 13276 / CIP 106301)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
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)
Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Shewanella oneidensis (strain MR-1)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10090
-
90-MtCM, MALDI-TOF mass spectrometry and calculated from amino acid sequence
11300
-
subunit, calculated from amino acid sequence
11700
the recombinant protein AroQMs in Escherichia coli
11770
-
105-MtCM, MALDI-TOF mass spectrometry and calculated from amino acid sequence
17800
in Escherichia coli for the mature *AroQMs protein
18540
-
subunit, calculated from amino acid sequence
18600
-
monomer, calculated, 167-residue untagged lederless MtCM; monomer, mass spectrometry, 167-residue untagged lederless MtCM
19540
-
monomer, calculated from the variant that lacks 33 N-terminal residues; monomer, mass spectrometry
19670
-
monomer, calculated from the leaderless variant with additional start methionine; monomer, mass, spectrometry, leaderless variant with additional start methionine
22000
-
monomer, SDS-PAGE
23000
-
monomer, SDS-PAGE, 167-residue untagged lederless MtCM
23500
-
gel filtration
25790
-
gel filtration
27900
calculated from amino acid sequence
29390
-
gel filtration
30110
predicted from amino acid sequence
35400
-
ultracentrifugation
36000
-
enzyme form CM2, gel filtration
37000
-
monomeric molecular mass 18474 Da. Absorbance, liquid chromatography-mass spectrometry
40000
-
gel filtration
43400
-
gel filtration
45000
-
enzyme form CM2, gel filtration
47590
gel filtration
48000
-
enzyme form CM2, gel filtration
51000
-
sucrose density gradient centrifugation
52000
-
enzyme form CM1, gel filtration
59000
-
gel filtration, another enzyme form with MW 55000 exists
60000
Claviceps sp.
-
gel filtration
61000
-
gel filtration
62200
gel filtration
63000
-
gel filtration
75000
-
enzyme form CM1, gel filtration
78000
-
sedimentation equilibrium analysis
91000
-
gel filtration, in presence of Phe or Tyr a MW of 175000 is measured
140000
-
chorismate mutase/prephenate dehydratase, enzyme form CM2, gel filtration
160000 - 180000
-
disc gel electrophoresis
187000
-
chorismate mutase/prephenate dehydratase, gel filtration
320000
-
chorismate mutase/prephenate dehydratase, gel filtration
420000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
-
x-ray crystallography
homodimer
homohexamer
monomer
tetramer
-
4 * 47000, chorismate mutase/prephenate dehydratase, SDS-PAGE
trimer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures of double mutants C88S/R90K and C88K/R90S, hanging drop vapour diffusion method at room temperature, space group R3 with a and b: 82.6 A and c: 42.8 A
-
hanging drop vapor-diffusion method at room temperature and high ionic strength, orthorhombic space group P212121 with a: 52.2 A, b: 83.8 A, c: 86.0 A, nine sulfate ions, five glycerol molecules, 424 water molecules
-
Performance of molecular dynamics simulations for the three enzyme-ligand complexes(CHOR,PRE and TSA) in addition to the TPS calculations. 8-hydroxy-2-oxa-bicyclo[3.3.1]non-6-ene-3,5-dicarboxylic acid as a TSA. The principal component analysis (PCA) to analyze structures is used
*MtCM, encoded by ORF Rv1885c in strain H37Rv. First characterized example of an AroQgamma fold. Description of the crystal optimization of a protein target that crystallizes very rapidly. 1. 175-residue version of *MtCM (encoded on plasmid pKTU3-HCT, dissolved in 20 mM potassium phosphate buffer pH7.5. 2. leaderless untagged 167-residue version of *MtCM encoded by plasmid pKTU3-HT used, buffered with 20 mM Tris-HCl pH 8.0)
37.2 kDa. Each asymmetric unit contains a homodimer, corresponding to the two protomers of the biological dimer. *MtCM as a model system for the *AroQ subclass and determined its crystal structure at high resolution, both in its unliganded form and in complex with transition state analog (1S,3S,5R,6R)-6-hydroxy-4-oxabicyco[3.3.1]non-7-ene-1,3-dicarboxylic acid. Heavy-atom derivatives prepared. Successful heavy-atom compounds are lead (II) acetate and thallium (III) acetate
-
alone and in complex with 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase, hanging drop vapor diffusion method, using 25% (w/v) PEG 1500 and 0.1 M MMT buffer (L-malic acid, MES, Tris) pH 8.0-9.0
Analysis of the structure shows a novel fold topology for the protein with a topologically rearranged helix containing R134. *MtCM does not have an allosteric regulation site
-
analysis reveals the presence of two monomers in the asymmetric unit
-
in complex with L-malate, hanging drop vapor diffusion method, using 15% (w/v) PEG 1500 and 0.1 M of the L-malate-containing MMT buffer system pH 8.0-9.0
-
MtCM (Rv1885c) (PDB ID-2F6L), docked conformation of (3S,6Z)-8-hydroxy-2-oxabicyclo[3.3.3]undec-6-ene-3,5-dicarboxylic acid, 4-[[2-(3,4-dimethoxyphenyl)ethyl]amino]-3-nitro-5-sulfamoylbenzoic acid, and (3S,6Z)-8-hydroxy-2-azabicyclo[3.3.3]undec-6-ene-3,5-dicarboxylic acid, (2Z)-2-(4-chlorophenyl)-3-(4,5-dimethoxy-2-nitrophenyl)prop-2-enoic acid in the catalytic site of MtCM x-ray crystal structure is shown
-
of the homodimeric chorismate mutase (Rv1885c). The crystal structure corresponds to the AroQ class CM of Mycobacterium tuberculosis. Determination of the crystal structure of the unique extracytoplasmic MtbCM in complex with its allosteric ligand, L-Trp. Se-Met MtbCM crystallizes in space group C2 in the presence of Trp. The Mycobacterium tuberculosis enzyme is an all-helical protein. Structural comparisons show that CMs from different organisms have envolved into two completely unrelated protein folds, suggesting separate evolutionary origins of the enzyme. On the basis of the structural fold adopted by the protein, CMs have been classified into the AroH and AroQ classes
-
sitting drop vapour diffusion method, using 0.1 M Tris-HCl (pH 8.6), 0.2 M MgCl2, and 20% poly(ethylene glycol) 400 for the 90 amino acid enzyme 90-MtCM
-
crystal structure of the T state of the allosteric enzyme and comparison with the R state
-
crystal structure of wild-type enzyme cocrystallized with Trp and an endo-exabicyclic transition state analogue inhibitor, of wild-type enzyme cocrystallized with Tyr and the endo-oxabicyclic transition state analogue inhibitor and of the Thr226Ser mutant enzyme in complex with Trp
-
Thr226Ile mutant enzyme
-
The corresponding positional fluctuations from the MD simulation are in good agreement with those obtained by X-ray crystallography
-
hanging drop vapour diffusion method, in 2 M ammonium sulfate, 0.1 M citrate/phosphate buffer, pH 4.2
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
heat inactivation above
42
-
4 min, 59% loss of chorismate mutase P activity, no effect on chorismate mutase T activity
50 - 60
-
5 min, enzyme form CM2, stable
50
half-life of 42.7 min
52
-
20 min, 50% loss of activity, enzyme form CM1; 5 min, more than 50% loss of activity, enzyme form CM2
54
-
1 h, 50% loss of activity, wild-type enzyme
55
-
pH 9.0, 5 min, enzyme form CM2, 15% loss of activity
58
-
1 h, 50% inactivation, genetically engineered enzyme with amino acid residues 1-285
62
-
1 h, 50% inactivation, genetically engineered enzyme with amino acid residues 1-300
63
-
midpoint of unfolding transitions
70
half-life of 8.1 min
80
half-life of 6.5 min
88
-
midpoint of unfolding transitions
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
chorismate mutase irreversibly immobilized by n-decylamine-substituted agarose allows repeated quantitative conversion of chorismate to prephenate
-
the enzyme is stable in high concentrations of glycerol, above 10% v/v, pH 7.5-8.0, and in the presence of thiols, citrate, and NAD+
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, enzyme form CM1 and CM2 are stable
-
-20C, enzyme form CM2, less than 10% loss of activity after 6 months
-
-20C, partially purified enzyme, stable for 7 days
-
-25C, in presence of PMSF, stable for at least 2 months
-
-78C, 100 mM N-ethylmorpholine, 1.0 mM dithioerythritol, 1.0 mM N-ethylene-diaminetetraacetic acid, 21 mM trisodium citrate, 10% (v/v) glycerol in doubly distilled water adjusted to pH 7.0 with concentrated HCl
-
0C or -20C, enzyme form CM1, stable for at least 1 month
-
0C or -20C, enzyme form CM2, stable for at least 2 weeks
-
0C or -20C, enzyme forms CM1 and CM2, stable for at least 6 weeks
-
4C, 0.1 mM Trp, stable for 5 days with a rapid loss in activity after this time
-
4C, in presence of PMSF, stable for at least 1 month
-
4C, stable for 1 month
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
*MtCM, which has a melting temperature of 48C, rapidly renatures after heat denaturation, and have subsequently exploited this feature for purifying the enzyme
-
ammonium sulfate precipitation, Ni-NTA agarose column chromatography, and Superdex 75 gel filtration
anion exchange column chromatography, Superdex 75 gel filtration
-
chorismate mutase/prephenate dehydratase
-
enzyme form CM1 and CM2
gel filtration
His-Bind protein purification column chromatography
-
in association with prephenate dehydratase
-
N-terminal Met1 is in most samples removed
-
Ni-NTA column chromatography
-
Ni2+ affinity column chromatography and gel filtration
-
Ni2+-affinity column chromatograp