BRENDA - Enzyme Database
show all sequences of 1.13.11.11

Structural and functional analyses of human tryptophan 2,3-dioxygenase

Meng, B.; Wu, D.; Gu, J.; Ouyang, S.; Ding, W.; Liu, Z.; Proteins 82, 3210-3216 (2014)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3)
Homo sapiens
Crystallization (Commentary)
Crystallization
Organism
purified full-length and truncated enzyme variants without heme cofactor, hanging drop vapor diffusion method, mixing of 0.001 ml of 20 mg/ml protein in 100 mM Tris–HCl, pH 8.0, with 0.001 ml of reservoir solution containing 0.1 M sodium cacodylate, pH 6.0-6.5, 6-10% w/v MPEG 5000, and 10% v/v 2-propanol, addition of hexammine cobalt(III) chloride improves the diffraction significantly, 16°C, X-ray diffraction structure determination and analysis at 2.90 A resolution
Homo sapiens
Engineering
Amino acid exchange
Commentary
Organism
F140A
site-directed mutagenesis, the mutant shows 0.25% of wild-type activity
Homo sapiens
F72A
site-directed mutagenesis, inactive mutant
Homo sapiens
H328A
site-directed mutagenesis, inactive mutant
Homo sapiens
H76A,
site-directed mutagenesis, inactive mutant
Homo sapiens
additional information
full-length hTDO is unstable, and various truncations are constructed. A truncation containing amino acids 19-388 is found to have good solubility and stability, the truncated mutant shows 80.45% of wild-type activity
Homo sapiens
R144A
site-directed mutagenesis, the mutant shows 0.88% of wild-type activity
Homo sapiens
S151A
site-directed mutagenesis, the mutant shows 9.08% of wild-type activity
Homo sapiens
Y42A
site-directed mutagenesis, the mutant shows 0.5% of wild-type activity
Homo sapiens
Y45A
site-directed mutagenesis, the mutant shows 1.13% of wild-type activity
Homo sapiens
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten kinetics
Homo sapiens
0.135
-
L-tryptophan
pH 8.0, 25°C, recombinant wild-type enzyme
Homo sapiens
0.146
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant S151A
Homo sapiens
0.153
-
L-tryptophan
pH 8.0, 25°C, recombinant truncated enzyme mutant
Homo sapiens
0.227
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y42A
Homo sapiens
0.631
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant R144A
Homo sapiens
0.661
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y45A
Homo sapiens
1.531
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant F140A
Homo sapiens
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
L-tryptophan + O2
Homo sapiens
-
N-formyl-L-kynurenine
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Homo sapiens
P48775
-
-
Purification (Commentary)
Commentary
Organism
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli BL21(DE3) by nickel affinity chromatography and gel filtration
Homo sapiens
Source Tissue
Source Tissue
Commentary
Organism
Textmining
liver
mainly
Homo sapiens
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
L-tryptophan + O2
-
743760
Homo sapiens
N-formyl-L-kynurenine
-
-
-
?
L-tryptophan + O2
cleavage of the C2-C3 bond in the indole moiety of L-Trp and incorporation of one oxygen molecule
743760
Homo sapiens
N-formyl-L-kynurenine
-
-
-
?
additional information
enzyme TDO is highly substrate-specific for L-Trp and the related derivatives 6-fluoro-Trp and 5-fluoro-Trp
743760
Homo sapiens
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
dimer
apo hTDO
Homo sapiens
More
hTDO must form an oligomer to exhibit activity. Active site structure, overview
Homo sapiens
tetramer
holo-hTDO, dimer of dimers, in which the two C-shape dimers (AB and CD) are clamped perpendicularly to each other to form a tight tetramer
Homo sapiens
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Homo sapiens
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.01
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y42A
Homo sapiens
0.032
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant F140A
Homo sapiens
0.045
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant R144A
Homo sapiens
0.062
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y45A
Homo sapiens
0.105
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant S151A
Homo sapiens
0.976
-
L-tryptophan
pH 8.0, 25°C, recombinant truncated enzyme mutant
Homo sapiens
1071
-
L-tryptophan
pH 8.0, 25°C, recombinant wild-type enzyme
Homo sapiens
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Homo sapiens
Cofactor
Cofactor
Commentary
Organism
Structure
heme
binding structure, overview
Homo sapiens
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3)
Homo sapiens
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
heme
binding structure, overview
Homo sapiens
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified full-length and truncated enzyme variants without heme cofactor, hanging drop vapor diffusion method, mixing of 0.001 ml of 20 mg/ml protein in 100 mM Tris–HCl, pH 8.0, with 0.001 ml of reservoir solution containing 0.1 M sodium cacodylate, pH 6.0-6.5, 6-10% w/v MPEG 5000, and 10% v/v 2-propanol, addition of hexammine cobalt(III) chloride improves the diffraction significantly, 16°C, X-ray diffraction structure determination and analysis at 2.90 A resolution
Homo sapiens
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
F140A
site-directed mutagenesis, the mutant shows 0.25% of wild-type activity
Homo sapiens
F72A
site-directed mutagenesis, inactive mutant
Homo sapiens
H328A
site-directed mutagenesis, inactive mutant
Homo sapiens
H76A,
site-directed mutagenesis, inactive mutant
Homo sapiens
additional information
full-length hTDO is unstable, and various truncations are constructed. A truncation containing amino acids 19-388 is found to have good solubility and stability, the truncated mutant shows 80.45% of wild-type activity
Homo sapiens
R144A
site-directed mutagenesis, the mutant shows 0.88% of wild-type activity
Homo sapiens
S151A
site-directed mutagenesis, the mutant shows 9.08% of wild-type activity
Homo sapiens
Y42A
site-directed mutagenesis, the mutant shows 0.5% of wild-type activity
Homo sapiens
Y45A
site-directed mutagenesis, the mutant shows 1.13% of wild-type activity
Homo sapiens
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
Michaelis-Menten kinetics
Homo sapiens
0.135
-
L-tryptophan
pH 8.0, 25°C, recombinant wild-type enzyme
Homo sapiens
0.146
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant S151A
Homo sapiens
0.153
-
L-tryptophan
pH 8.0, 25°C, recombinant truncated enzyme mutant
Homo sapiens
0.227
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y42A
Homo sapiens
0.631
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant R144A
Homo sapiens
0.661
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y45A
Homo sapiens
1.531
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant F140A
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
L-tryptophan + O2
Homo sapiens
-
N-formyl-L-kynurenine
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli BL21(DE3) by nickel affinity chromatography and gel filtration
Homo sapiens
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
liver
mainly
Homo sapiens
-
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
L-tryptophan + O2
-
743760
Homo sapiens
N-formyl-L-kynurenine
-
-
-
?
L-tryptophan + O2
cleavage of the C2-C3 bond in the indole moiety of L-Trp and incorporation of one oxygen molecule
743760
Homo sapiens
N-formyl-L-kynurenine
-
-
-
?
additional information
enzyme TDO is highly substrate-specific for L-Trp and the related derivatives 6-fluoro-Trp and 5-fluoro-Trp
743760
Homo sapiens
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
apo hTDO
Homo sapiens
More
hTDO must form an oligomer to exhibit activity. Active site structure, overview
Homo sapiens
tetramer
holo-hTDO, dimer of dimers, in which the two C-shape dimers (AB and CD) are clamped perpendicularly to each other to form a tight tetramer
Homo sapiens
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
25
-
assay at
Homo sapiens
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.01
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y42A
Homo sapiens
0.032
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant F140A
Homo sapiens
0.045
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant R144A
Homo sapiens
0.062
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant Y45A
Homo sapiens
0.105
-
L-tryptophan
pH 8.0, 25°C, recombinant mutant S151A
Homo sapiens
0.976
-
L-tryptophan
pH 8.0, 25°C, recombinant truncated enzyme mutant
Homo sapiens
1071
-
L-tryptophan
pH 8.0, 25°C, recombinant wild-type enzyme
Homo sapiens
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8
-
assay at
Homo sapiens
General Information
General Information
Commentary
Organism
additional information
eight residues play critical roles in L-tryptophan oxidation, i.e. Y42, Y45, F72, H76, F140, R144, S151, and H328. hTDO must form an oligomer to exhibit activity
Homo sapiens
physiological function
tryptophan 2,3-dioxygenase (TDO) is one of the two key enzymes in the kynurenine pathway, it catalyzes the indole ring cleavage at the C2-C3 bond of L-tryptophan. This is a rate-limiting step in the regulation of tryptophan concentration in vivo. In addition to its role in protein synthesis, 95% of L-Trp in the human body is processed by the kynurenine pathway, leading to the production of nicotinamide adenine dinucleotide. Enzyme TDO is expressed in many tumor cells and is related to reduction of antitumor immune response
Homo sapiens
General Information (protein specific)
General Information
Commentary
Organism
additional information
eight residues play critical roles in L-tryptophan oxidation, i.e. Y42, Y45, F72, H76, F140, R144, S151, and H328. hTDO must form an oligomer to exhibit activity
Homo sapiens
physiological function
tryptophan 2,3-dioxygenase (TDO) is one of the two key enzymes in the kynurenine pathway, it catalyzes the indole ring cleavage at the C2-C3 bond of L-tryptophan. This is a rate-limiting step in the regulation of tryptophan concentration in vivo. In addition to its role in protein synthesis, 95% of L-Trp in the human body is processed by the kynurenine pathway, leading to the production of nicotinamide adenine dinucleotide. Enzyme TDO is expressed in many tumor cells and is related to reduction of antitumor immune response
Homo sapiens
Other publictions for EC 1.13.11.11
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
743153
Nienhaus
Substrate binding primes huma ...
Homo sapiens
J. Phys. Chem. B
121
7412-7420
2017
-
-
-
-
-
-
-
1
-
1
-
1
-
3
-
-
-
1
-
-
-
-
2
-
-
-
-
-
1
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
2
2
-
-
-
743358
Gonzalez Esquivel
Kynurenine pathway metabolite ...
Homo sapiens, Mus musculus, Xanthomonas campestris pv. campestris, Xanthomonas campestris pv. campestris ATCC 33913
Neuropharmacology
112
331-345
2017
2
-
-
-
-
-
3
2
-
-
-
-
-
7
-
-
-
-
-
29
-
-
4
2
-
-
-
-
-
-
-
3
-
-
-
2
-
-
5
-
-
-
-
3
-
3
-
-
-
-
-
-
-
-
-
31
-
-
4
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
741938
Basran
Analysis of reaction intermed ...
Homo sapiens, Xanthomonas campestris pv. campestris, Xanthomonas campestris pv. campestris ATCC 33913
Biochemistry
55
6743-6750
2016
-
-
2
-
-
-
-
2
-
2
-
3
-
4
-
-
2
-
-
-
-
-
12
-
2
-
-
-
2
-
-
2
-
-
-
-
-
2
2
-
-
-
-
-
-
2
-
2
-
3
-
-
-
2
-
-
-
-
12
-
2
-
-
-
2
-
-
-
-
4
4
-
-
-
742986
Pantouris
Insights into the mechanism o ...
Homo sapiens
J. Enzyme Inhib. Med. Chem.
31
70-78
2016
-
1
-
-
-
-
2
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
743821
Lewis-Ballester
Molecular basis for catalysis ...
Homo sapiens
Sci. Rep.
6
35169
2016
-
-
1
1
1
-
-
1
-
1
-
1
-
2
-
-
-
1
-
1
-
-
2
1
1
-
-
-
1
-
-
1
-
-
-
-
-
1
1
1
1
-
-
-
-
1
-
1
-
1
-
-
-
-
-
1
-
-
2
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
743005
Yuasa
Efficient tryptophan-cataboli ...
Branchiostoma floridae, Caenorhabditis elegans, Drosophila melanogaster, Homo sapiens, Monosiga brevicollis, Nematostella vectensis, no activity in Brugia malayi, no activity in Saccharomyces cerevisiae, no activity in Schistosoma mansoni, Rattus norvegicus, Strongylocentrotus purpuratus
J. Exp. Zool. B
324
128-140
2015
-
-
8
-
-
-
-
11
-
-
-
8
-
14
-
-
-
-
-
-
-
-
8
-
8
-
-
-
3
-
-
1
-
-
-
-
-
8
1
-
-
-
-
-
-
11
-
-
-
8
-
-
-
-
-
-
-
-
8
-
8
-
-
-
3
-
-
-
-
8
8
-
-
-
743064
Wu
Identification of substituted ...
Homo sapiens
J. Med. Chem.
58
7807-7819
2015
-
1
1
-
-
-
21
-
-
-
-
1
-
1
-
-
1
-
-
2
-
-
1
-
1
-
-
-
1
-
-
1
-
-
20
-
1
1
1
-
-
-
20
21
-
-
-
-
-
1
-
-
-
1
-
2
-
-
1
-
1
-
-
-
1
-
-
-
-
3
3
-
-
-
742114
Maeta
Contributions of tryptophan 2 ...
Mus musculus, Mus musculus C57BL/6
Biosci. Biotechnol. Biochem.
78
878-881
2014
-
-
-
-
1
-
-
-
-
-
-
6
-
3
-
-
-
-
-
1
-
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
6
-
-
-
-
-
1
-
-
8
-
-
-
-
-
-
-
-
-
-
3
3
-
-
-
743760
Meng
Structural and functional ana ...
Homo sapiens
Proteins
82
3210-3216
2014
-
-
1
1
9
-
-
8
-
-
-
1
-
2
-
-
1
-
-
1
-
-
3
3
1
-
-
7
1
-
-
1
-
-
-
-
-
1
1
1
9
-
-
-
-
8
-
-
-
1
-
-
-
1
-
1
-
-
3
3
1
-
-
7
1
-
-
-
-
2
2
-
-
-