Information on EC 4.1.1.48 - indole-3-glycerol-phosphate synthase

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

EC NUMBER
COMMENTARY hide
4.1.1.48
-
RECOMMENDED NAME
GeneOntology No.
indole-3-glycerol-phosphate synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate = 1-C-(indol-3-yl)glycerol 3-phosphate + CO2 + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
decarboxylation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
L-tryptophan biosynthesis
-
-
Metabolic pathways
-
-
Phenylalanine, tyrosine and tryptophan biosynthesis
-
-
tryptophan metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate carboxy-lyase [cyclizing; 1-C-(indol-3-yl)glycerol-3-phosphate-forming]
In some organisms, this enzyme is part of a multifunctional protein, together with one or more other components of the system for the biosynthesis of tryptophan [EC 2.4.2.18 (anthranilate phosphoribosyltransferase), EC 4.1.3.27 (anthranilate synthase), EC 4.2.1.20 (tryptophan synthase) and EC 5.3.1.24 (phosphoribosylanthranilate isomerase)].
CAS REGISTRY NUMBER
COMMENTARY hide
9031-60-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
possibly forms a bifunctional enzyme with phosphoribosylanthranilate isomerase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
Enterobacter liquefaciens
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
strain W3110
-
-
Manually annotated by BRENDA team
multifunctional enzyme with the activities of Ec 4.1.4.27, Ec 4.2.2.18, Ec 4.1.1.48 and Ec 4.2.1.20
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, Ec4.1.1.48/Ec5.3.1.24
-
-
Manually annotated by BRENDA team
wild-type strain 655 and mutant strains
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Vitis vinifera x Vitis riparia
-
-
-
Manually annotated by BRENDA team
Vitis vinifera x Vitis vinifera
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
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
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
1-(3-indolyl)glycerol-3-phosphate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
1-C-(indol-3-yl)glycerol 3-phosphate + CO2 + H2O
show the reaction diagram
1-(2-Carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
1-(3-Indolyl)glycerol-3-phosphate + CO2 + H2O
show the reaction diagram
1-(2-Carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
?
show the reaction diagram
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
indoleglycerol phosphate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate + ?
1-C-(indol-3-yl)glycerol 3-phosphate + CO2 + H2O
show the reaction diagram
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
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
1-(3-indolyl)glycerol-3-phosphate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
1-C-(indol-3-yl)glycerol 3-phosphate + CO2 + H2O
show the reaction diagram
1-(2-Carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
1-(3-Indolyl)glycerol-3-phosphate + CO2 + H2O
show the reaction diagram
-
-
-
ir
1-(2-Carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
?
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
significantly affects activity, the optimal concentration is about 0.4-2.0 mM
Mg2+
-
significantly affects activity, the optimal concentration is about 0.4-2.0 mM
Mn2+
-
significantly affects activity, the optimal concentration is about 0.4-2.0 mM
Na+
-
significantly affects activity, the optimal concentration is about 0.4-2.0 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ATB107
geomycine
-
0.5 mg/ml
H2O2
-
0.001% H2O2 inactivates the enzyme almost completely
kanamycin
-
0.5 mg/ml
L-tryptophan
tryptophan at high concentrations has definite inhibitory activity against Mycobacterium tuberculosis but does not antagonize the activity of ATB107
methanol
-
enzyme activity is gradually lost in the presence of increasing concentrations of methanol and completely lost at 65%
phosphate
-
-
Reduced 1-[2-carboxyphenyl)amino]-1-deoxyribulose 5-phosphate
-
-
-
streptomycin
-
0.5 mg/ml
tryptophan
tryptophan at high concentrations has definite inhibitory activity against Mycobacterium tuberculosis but does not antagonize the activity of ATB107
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000013 - 0.09
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
0.000006 - 21.67
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 11.1
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
0.025 - 7.2
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.36 - 16000
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate
100 - 14200
1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.27
phosphate
-
60C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.004
-
recombinant enzyme, at 37C, pH 7.0
additional information
-
rapid and sensitive continous spectrophotofluorometric assay that can be used in presence of glycerol
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
in 5 mM Tris-HCl buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
-
in 5 mM Tris-HCl buffer
7 - 8.2
-
activity begins to decrease above pH 8.2 and is 70% of maximum at pH 9.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 35
-
activation energy value of 8.4 kcal/mol, the linearity of the Arrhenius plot demonstrates that there is no change in the rate-limiting step in the temperature range employed
60 - 95
60C: about 50% of maximal activity, 95C: about 45% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Brucella abortus (strain 2308)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
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)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
21000
-
gel filtration
22000
-
gel filtration
23500
-
gel filtration, N-terminal deletion mutant lacking 26 amino acids
23600
-
gel filtration
24200
gel filtration, N-terminal deletion mutant lacking 25 amino acids
26300
-
gel filtration, wild-type
28000
-
SDS-PAGE, recombinant enzyme
28200
gel filtration, wild-type
30000
-
gel-filtration chromatography
32000
-
gel filtration
39300
gel filtration, N-terminal deletion mutant lacking 25 amino acids
44000
-
equilibrium sedimentation
47100
gel filtration, wild-type
320000 - 340000
-
anthranilate synthetase complex, gel filtration
325000
-
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
tetramer
-
2 * 94000 + 2 * 70000, anthranilate synthetase complex, one subunit is a trifunctional peptide which contains the catalytic sites for the phosphoribosylanthranilate isomerase and indoleglycerol phosphate synthetase reactions, and associates with the second subunit to form glutamine-dependent anthranilate synthetase, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
kinetic studies of folding mechanism
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2.0 A structure; vapour diffusion method in hanging drops, the 2.0 A crystal structure is determined and compared with the known 2.0 A structure of the IGPS domain of the bifunctional enzyme from Escherichia coli. Both enzymes have only 30% sequence identity, but share high structural similarity
-
crystal packing seems to be influenced by ionic strength of the solvent
-
in complex with substrate 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate, with a substrate analogue, and with product indole-3-glycerol phosphate
-
mutagenesis data and crystal structure analysis of IGPS from Sulfolobus solfataricus allows for the formulation of a plausible chemical mechanism of the reaction
-
N-terminal deletion mutant lacking 26 amino acids, structure of core is unchanged compared to wild-type
-
contains 17 strong salt bridges
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
slight activity at pH 5.0
678351
8
-
almost no activity at pH 8.0
678351
additional information
-
melting temperature is higher at alkaline that at neutral pH
4610
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 50
-
little loss of activity after a 10 min heating in a water bath at temperatures from 40 to 50C
44
-
half-life: 1.8 min
73
-
335 min, half-life for irreversible thermal inactivation
83.5
-
110 min, half-life for irreversible thermal inactivation
84
-
110 min, half-life for irreversible thermal denaturation
85.5
-
14.4 min, half-life for irreversible thermal inactivation, mutant R241A; 36.1 min, half-life for irreversible thermal inactivation, mutant D184A; 42.3 min, half-life for irreversible thermal inactivation
86.5
-
in 0.05 M potassium phosphate at pH 7.5. Half-life of wild-type enzyme: 46 min, half-life of mutant enzyme P2S: 18 min, hal-life of mutant enzyme F246S 1 min, half-life of mutant enzyme G212E: 38 min, half-life of mutant enzyme P2S/F246S: less than 0.1 min, half-life of mutant enzyme P2S/G212E: 19 min
89
-
half-life: 4.4 min
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
guanidinium-induced denaturation. Unfolding mechanism closely approaches a two-state model at pH 7.0 and a more complex mechanism at pH 9.0
-
half-life after trypsinolysis in 0.1 M Tris acetate at pH 7.8 and 25C: wild-type enzyme (120 min), mutant enzyme P2S (60 min), mutant enzyme F246S (40 min), mutant enzyme G212E (40 min), mutant enzyme P2S/F246S (8 min), mutant enzyme P2S/G212E (15 min)
-
optimal stabilization by 0.8 M sucrose
-
the enzyme is strongly stabilized in phosphate buffer (t1/2: 46 min at 87C) in comparison to HEPPS buffer (t1/2: 4.4 min at 89C)
-
the higher stability of the enzyme from Sulfolobus solfataricus compared with the enzyme from E. coli seems to be the result of several improved interactions. Including a large number of salt bridges, stabilization of alpha-helices and strengthening of both polypeptide chain termini and solvent-exposed loops
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
urea
-
the structure of partially folded states of the enzyme is assessed by hydrogen exchange mass spectrometry and G model simulations. HX-MS analysis of the peptic peptides derived from the pulse-labeled product of the submillisecond folding reaction from the urea-denatured state reveal strong protection in the (betaalpha)4 region, modest protection in the neighboring (betaalpha)13 and (betaalpha)5beta6 segments and no significant protection in the remaining N and C-terminal segments. The results demonstrate that this species is not a collapsed form of the unfolded state under native-favoring conditions nor is it the native state formed via fast-track folding
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, 5 mM Tris-HCl (pH 7.0), at least 2 months, remains stable
-
4C, 1 mM potassium phosphate buffer pH 7.5, 1 mM dithiothreitol, stable for hours
-
4C, in presence of 0.8 M sucrose, 20% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
bifunctional enzyme phosphoribosylanthranilate isomerase-indoleglycerol phosphate synthetase
chromatographic purification, Phenyl Sepharose High Sub FF column, ultrafiltration membrane (molecular weight cutoff) of 10,000 Da, Sephacryl S-100 column
-
final step with Superdex-75 gel filtration
-
monofunctional enzyme
-
multifunctional enzyme with the activities of Ec 4.1.4.27, Ec 4.2.2.18, Ec 4.1.1.48 and Ec 4.2.1.20
-
Ni-NTA His-binding resin affinity chromatography
-
Q-Sepharose column chromatography
-
TALON metal affinity resin column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli strain BL21
expressed in Escherichia coli strain BL21 (DE3)
-
expressed in Escherichia coli strain KK-8(pDM)
-
expression in Escherichia coli
expression in Escherichia coli BL21(DE3) STAR
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
a significant increase in trp mRNA level is observed in cells grown in medium depleted of L-tryptophan, compared to cells grown in medium supplemented with L-tryptophan, indicating that expression of the gene cluster is regulated at the transcriptional level
ATB107 does not affect the expression of IGPS
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L55S
-
Lys55Ser with 60fold decrease in turnover-number and 450fold increase in Km-value
L5V
-
Leu5Val without effect on turnover-number and Km
E168A
site directed mutagenesis
H170A
site directed mutagenesis
L193A
site directed mutagenesis
L196A
site directed mutagenesis
N189A
site directed mutagenesis
N192A
site directed mutagenesis
P63A
site directed mutagenesis
R191A
site directed mutagenesis
S64A
site directed mutagenesis
V169A
site directed mutagenesis
E168A
site directed mutagenesis
H170A
site directed mutagenesis
L193A
site directed mutagenesis
L196A
site directed mutagenesis
N189A
site directed mutagenesis
N192A
site directed mutagenesis
P63A
site directed mutagenesis
R191A
site directed mutagenesis
S64A
site directed mutagenesis
V169A
site directed mutagenesis
E168A
-
site directed mutagenesis
-
L196A
-
site directed mutagenesis
-
R191A
-
site directed mutagenesis
-
S64A
-
site directed mutagenesis
-
D61C
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is about 1.5fold lower than the wild-type value
E210Q
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 6fold lower than wild-type value
E51Q
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 28fold lower than wild-type value
F246S
-
about 2fold increase in turnover-number, 8fold increase in KM-value, decrease in kcat/Km
F89A
-
approximately 24fold increase in the KM-value, eleven-fold decrease in the maximum turnover rate
G212E
-
about 2fold increase in turnover-number, 100fold increase Km-value, decrease in kcat/Km. Mutant enzyme is about as thermostable as wild-type enzyme
K53Q
-
low activity, kcat is 570fold lower than wild-type value
K53R
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 500fold lower than wild-type value
M237T
-
kcat/Km of 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate is fold than wild-type value
N90A
-
loss of the descending limb in the pH rate profile, 5-fold decrease in kcat at 25C, but at higher temperatures (i.e. 75C), the kinetic parameters for the N90A mutant enzyme closer to wild-type enzyme. Mutant enzyme has substantially reduced solvent deuterium kinetic isotope effects compared to wild-type enzyme at 75C. 2fold decrease in the rate of thermal inactivation at 90C as compared to wild-type enzyme
N90Q
-
the steady-state kinetics for the mutant enzyme are substantially decreased compared to wild-type enzyme at lower temperatures, but kcat of the N90Q variant approaches that of wild-type enzyme at higher temperatures. The solvent deuterium kinetic isotope effects on kcat is also substantially reduced compared to wild-type enzyme. 2fold decrease in the rate of thermal inactivation at 90C
P2S
-
slight increase in kcat/Km (substrate: 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose-5-phosphate). Mutant enzyme is about as thermostable as wild-type enzyme
P2S/F246S
-
about 2fold increase in turnover-number, 8fold increase in KM-value, decrease in kcat/Km
P2S/G212E
-
about 4fold increase in turnover-number,200fold increase in Km-valuer, decrease in kcat/Km
R182A
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 39fold lower than wild-type value
R18C
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is about 1.5fold lower than the wild-type value
R54A
-
mutation does not result in any substantial change to the steady-state kinetic parameters at any of the temperatures assayed (25C, 37C, 75C). At 75C, the mutant enzyme shows a small increase to kcat (about 1.8 fold) compared to wild-type enzyme. Mutant enzyme has substantially reduced solvent deuterium kinetic isotope effects compared to wild-type enzyme at 75C. Thermal inactivation constant at 90C is similar to wild-typ enzyme
R54A/N90A
-
the R54A/N90A double substitution are not additive with the effects of the R54A and N90A single substitutions suggesting some type of thermodynamic coupling between these residues
D61C
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is about 1.5fold lower than the wild-type value
-
E210Q
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 6fold lower than wild-type value
-
E51Q
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 28fold lower than wild-type value
-
K53Q
-
low activity, kcat is 570fold lower than wild-type value
-
K53R
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 500fold lower than wild-type value
-
R182A
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is 39fold lower than wild-type value
-
R18C
-
kcat/Km for 1-(2-carboxyphenylamino)-1-deoxy-D-ribulose 5-phosphate is about 1.5fold lower than the wild-type value
-
D184A
-
no alteration of native structure, very little difference in Km, turnover rate compared to wild type, but less thermal stability
R241A
-
no alteration of native structure, very little difference in Km, turnover rate compared to wild type, but less thermal stability
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
initiated by rapid 11fold dilution in refolding buffer (10 mM phosphate pH 7.8 and 25C)
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
-
indole-3-glycerol phosphate synthase of Mycobacterium tuberculosis is an attractive target for new anti-Tuberculosis drug development
medicine
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