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Literature summary for 1.13.11.11 extracted from

  • Meng, B.; Wu, D.; Gu, J.; Ouyang, S.; Ding, W.; Liu, Z.
    Structural and functional analyses of human tryptophan 2,3-dioxygenase (2014), Proteins, 82, 3210-3216 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli BL21(DE3) Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) 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

Protein Variants

Protein Variants Comment 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 Comment 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 Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-tryptophan + O2 Homo sapiens
-
N-formyl-L-kynurenine
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens P48775
-
-

Purification (Commentary)

Purification (Comment) 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 Comment Organism Textmining
liver mainly Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-tryptophan + O2
-
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 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 Homo sapiens ?
-
?

Subunits

Subunits Comment 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

Synonyms

Synonyms Comment Organism
hTDO
-
Homo sapiens
TDO
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment 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 Comment Organism
8
-
assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
heme binding structure, overview Homo sapiens

General Information

General Information Comment 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