US EPA

1529690 

Journal Article 

Complete Reaction Mechanism of Indoleamine 2,3-Dioxygenase as Revealed by QM/MM Simulations 

Capece, L; Lewis-Ballester, A; Yeh, SRu; Estrin, DA; Marti, MA 

2012 

Yes 

Journal of Physical Chemistry B
ISSN: 1520-6106
EISSN: 1520-5207 

116 

4 

1401-1413 

22196056 

10.1021/jp2082825 

WOS:000300461300022 

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 C-2=C-3 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-NH3+ to the epoxide oxygen, triggering epoxide ring
opening and a concerted nucleophilic attack of the ferryl oxygen to the C-2 of Tip that leads to
a metastable reaction intermediate. This intermediate subsequently converts to NFK, following C-
2-C-3 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.