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Literature summary for 1.13.11.11 extracted from

  • Lewis-Ballester, A.; Forouhar, F.; Kim, S.M.; Lew, S.; Wang, Y.; Karkashon, S.; Seetharaman, J.; Batabyal, D.; Chiang, B.Y.; Hussain, M.; Correia, M.A.; Yeh, S.R.; Tong, L.
    Molecular basis for catalysis and substrate-mediated cellular stabilization of human tryptophan 2,3-dioxygenase (2016), Sci. Rep., 6, 35169 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of full-length His6-tagged hTDO protein in human liver HepG2 cell culture Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified enzyme in a ternary complex with the substrates L-Trp and O2 and in a binary complex with the product N-formylkynurenine, under-oil microbatch method under anaerobic conditions, mixing of 0.003 ml of 45 mg/ml protein in 50 mM Tris, pH 8.0, containing 150 mM NaCl and 10 mM L-Trp, and reduced with 2-fold molar excess of sodium dithionite, with 0.003-0.006 ml of precipitannt solution containing 50 mM sodium citrate, pH 5.6, 5% w/v PEG 3350, and 2% w/v Tacsimate, pH 5.0, X-ray diffraction structure determination and analysis at 2.5 A and 2.44 A resolution, respectively. Crystals of the Trp-bound binary complex are soaked in an O2-saturated precipitant solution supplemented with 20% v/v ethylene glycol at room temperature Homo sapiens

Protein Variants

Protein Variants Comment Organism
Y175G site-directed mutagenesis, the mutation leads to a 6fold slower multiple turnover velocity. In addition, pre-incubation of the Y175G mutant with 8 mM NFK retards the formation of the ternary complex by about 100fold and impedes Trp binding Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information stopped-flow kinetics Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ heme with Fe2+ coordinated by 4 N-atoms, overview 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
-
-

Reaction

Reaction Comment Organism Reaction ID
L-tryptophan + O2 = N-formyl-L-kynurenine the dioxygenation reaction is initiated by a direct attack of O2 on the C2 atom of the L-Trp indole ring, catalytic mechanism, overview. Exo binding site for L-Trp, located about 42 A from the active site and formed by residues conserved among tryptophan-auxotrophic TDOs. Trp binding at this exo site does not affect enzyme catalysis but instead it retards the degradation of hTDO through the ubiquitin-dependent proteasomal pathway Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
Hep-G2 cell
-
Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-tryptophan + O2
-
Homo sapiens N-formyl-L-kynurenine
-
?
additional information substrate recognition and binding structures, overview. The EG segment in hTDO plays a critical role in promoting NFK release, thereby allowing Trp binding during multiple turnover Homo sapiens ?
-
?

Subunits

Subunits Comment Organism
tetramer
-
Homo sapiens

Synonyms

Synonyms Comment Organism
TDO
-
Homo sapiens

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Homo sapiens

Cofactor

Cofactor Comment Organism Structure
heme with Fe2+ coordinated by 4 N-atoms, overview Homo sapiens

General Information

General Information Comment Organism
additional information the exo site regulates hTDO cellular stability Homo sapiens