Complete reaction mechanism of indoleamine 2,3-dioxygenase as revealed by QM/MM simulations

Luciana Capece, Ariel Lewis-Ballester, Syun-Ru Yeh, Dario A. Estrin, Marcelo A. Marti

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Indoleamine 2,3-dioxygenase (IDO) and tryptophan dioxygenase (TDO) are two heme proteins that catalyze the oxidation reaction of tryptophan (Trp) to N-formylkynurenine (NFK). Human IDO (hIDO) has recently been recognized as a potent anticancer drug target, a fact that triggered intense research on the reaction and inhibition mechanisms of hIDO. Our recent studies revealed that the dioxygenase reaction catalyzed by hIDO and TDO is initiated by addition of the ferric iron-bound superoxide to the C2=C3 bond of Trp to form a ferryl and Trp-epoxide intermediate, via a 2-indolenylperoxo radical transition state. The data demonstrate that the two atoms of dioxygen are inserted into the substrate in a stepwise fashion, challenging the paradigm of heme-based dioxygenase chemistry. In the current study, we used QM/MM methods to decipher the mechanism by which the second ferryl oxygen is inserted into the Trp-epoxide to form the NFK product in hIDO. Our results show that the most energetically favored pathway involves proton transfer from Trp-NH 3+ to the epoxide oxygen, triggering epoxide ring opening and a concerted nucleophilic attack of the ferryl oxygen to the C2 of Trp that leads to a metastable reaction intermediate. This intermediate subsequently converts to NFK, following C2-C3 bond cleavage and the associated back proton transfer from the oxygen to the amino group of Trp. A comparative study with Xantomonas campestris TDO (xcTDO) indicates that the reaction follows a similar pathway, although subtle differences distinguishing the two enzyme reactions are evident. The results underscore the importance of the NH3+ group of Trp in the two-step ferryl-based mechanism of hIDO and xcTDO, by acting as an acid catalyst to facilitate the epoxide ring-opening reaction and ferryl oxygen addition to the indole ring.

Original languageEnglish (US)
Pages (from-to)1401-1413
Number of pages13
JournalJournal of Physical Chemistry B
Volume116
Issue number4
DOIs
StatePublished - Feb 2 2012

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Indoleamine-Pyrrole 2,3,-Dioxygenase
tryptophan
Tryptophan
Oxygen
Dioxygenases
epoxy compounds
Epoxy Compounds
Proton transfer
simulation
Reaction intermediates
oxygen
Enzymes
rings
Iron
Atoms
Oxidation
Hemeproteins
Catalysts
Acids
reaction intermediates

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Materials Chemistry
  • Surfaces, Coatings and Films

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Complete reaction mechanism of indoleamine 2,3-dioxygenase as revealed by QM/MM simulations. / Capece, Luciana; Lewis-Ballester, Ariel; Yeh, Syun-Ru; Estrin, Dario A.; Marti, Marcelo A.

In: Journal of Physical Chemistry B, Vol. 116, No. 4, 02.02.2012, p. 1401-1413.

Research output: Contribution to journalArticle

Capece, Luciana ; Lewis-Ballester, Ariel ; Yeh, Syun-Ru ; Estrin, Dario A. ; Marti, Marcelo A. / Complete reaction mechanism of indoleamine 2,3-dioxygenase as revealed by QM/MM simulations. In: Journal of Physical Chemistry B. 2012 ; Vol. 116, No. 4. pp. 1401-1413.
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