Molecular basis for catalysis and substrate-mediated cellular stabilization of human tryptophan 2,3-dioxygenase

Ariel Lewis-Ballester, Farhad Forouhar, Sung Mi Kim, Scott Lew, Yongqiang Wang, Shay Karkashon, Jayaraman Seetharaman, Dipanwita Batabyal, Bing Yu Chiang, Munif Hussain, Maria Almira Correia, Syun-Ru Yeh, Liang Tong

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Abstract

Tryptophan 2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO) play a central role in tryptophan metabolism and are involved in many cellular and disease processes. Here we report the crystal structure of human TDO (hTDO) in a ternary complex with the substrates L-Trp and O2 and in a binary complex with the product N-formylkynurenine (NFK), defining for the first time the binding modes of both substrates and the product of this enzyme. The structure indicates that the dioxygenation reaction is initiated by a direct attack of O2 on the C2 atom of the L-Trp indole ring. The structure also reveals an exo binding site for L-Trp, located ∼42 Å from the active site and formed by residues conserved among tryptophan-auxotrophic TDOs. Biochemical and cellular studies indicate that 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. This exo site may therefore provide a novel L-Trp-mediated regulation mechanism for cellular degradation of hTDO, which may have important implications in human diseases.

Original languageEnglish (US)
Article number35169
JournalScientific Reports
Volume6
DOIs
StatePublished - Oct 20 2016

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Tryptophan Oxygenase
Catalysis
Tryptophan
Indoleamine-Pyrrole 2,3,-Dioxygenase
Enzymes
Ubiquitin
Catalytic Domain
Binding Sites

ASJC Scopus subject areas

  • General

Cite this

Lewis-Ballester, A., Forouhar, F., Kim, S. M., Lew, S., Wang, Y., Karkashon, S., ... Tong, L. (2016). Molecular basis for catalysis and substrate-mediated cellular stabilization of human tryptophan 2,3-dioxygenase. Scientific Reports, 6, [35169]. https://doi.org/10.1038/srep35169

Molecular basis for catalysis and substrate-mediated cellular stabilization of human tryptophan 2,3-dioxygenase. / Lewis-Ballester, Ariel; Forouhar, Farhad; Kim, Sung Mi; Lew, Scott; Wang, Yongqiang; Karkashon, Shay; Seetharaman, Jayaraman; Batabyal, Dipanwita; Chiang, Bing Yu; Hussain, Munif; Correia, Maria Almira; Yeh, Syun-Ru; Tong, Liang.

In: Scientific Reports, Vol. 6, 35169, 20.10.2016.

Research output: Contribution to journalArticle

Lewis-Ballester, A, Forouhar, F, Kim, SM, Lew, S, Wang, Y, Karkashon, S, Seetharaman, J, Batabyal, D, Chiang, BY, Hussain, M, Correia, MA, Yeh, S-R & Tong, L 2016, 'Molecular basis for catalysis and substrate-mediated cellular stabilization of human tryptophan 2,3-dioxygenase', Scientific Reports, vol. 6, 35169. https://doi.org/10.1038/srep35169
Lewis-Ballester, Ariel ; Forouhar, Farhad ; Kim, Sung Mi ; Lew, Scott ; Wang, Yongqiang ; Karkashon, Shay ; Seetharaman, Jayaraman ; Batabyal, Dipanwita ; Chiang, Bing Yu ; Hussain, Munif ; Correia, Maria Almira ; Yeh, Syun-Ru ; Tong, Liang. / Molecular basis for catalysis and substrate-mediated cellular stabilization of human tryptophan 2,3-dioxygenase. In: Scientific Reports. 2016 ; Vol. 6.
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