Nitration and inactivation of IDO by peroxynitrite

Hidetsugu Fujigaki, Kuniaki Saito, Felix Lin, Suwako Fujigaki, Kanako Takahashi, Brian M. Martin, Cai Y. Chen, Junichi Masuda, Jeffrey Kowalak, Osamu Takikawa, Mitsuru Seishima, Sanford P. Markey

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

IDO induction can deplete L-tryptophan in target cells, an effect partially responsible for the antimicrobial activities and antiallogeneic T cell responses of IFN-γ in human macrophages, dendritic cells, and bone marrow cells. L-Tryptophan depletion and NO production are both known to have an antimicrobial effect in macrophages, and the interaction of these two mechanisms is unclear. In this study we found that IDO activity was inhibited by the peroxynitrite generator, 3-(4-morpholinyl)sydnonimine, in PMA-differentiated cytokine-induced THP-1 (acute monocytic leukemia) cells and IFN-γ- stimulated PBMCs, whereas IDO protein expression was unaffected compared with that in untreated cells. Nitrotyrosine was detected in immunoprecipitated (IP)-IDO from PMA-differentiated cytokine-induced THP-1 cells treated with 3-(4-morpholinyl)sydnonimine, but not from untreated cells. Treatment of IP-IDO and recombinant IDO (rIDO) with peroxynitrite significantly decreased enzyme activity. Nitrotyrosine was detected in both peroxynitrite-treated IP-IDO and rIDO, but not in either untreated IP-IDO or rIDO. Peptide analysis by liquid chromatography/electrospray ionization and tandem mass spectrometry demonstrated that Tyr15, Tyr345, and Tyr353 in rIDO were nitrated by peroxynitrite. The levels of Tyr nitration and the inhibitory effect of peroxynitrite on IDO activity were significantly reduced in the Tyr 15-to-Phe mutant. These results indicate that IDO is nitrated and inactivated by peroxynitrite and that nitration of Tyr15 in IDO protein is the most important factor in the inactivation of IDO.

Original languageEnglish
Pages (from-to)372-379
Number of pages8
JournalJournal of Immunology
Volume176
Issue number1
DOIs
Publication statusPublished - 01-01-2006

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Fingerprint Dive into the research topics of 'Nitration and inactivation of IDO by peroxynitrite'. Together they form a unique fingerprint.

  • Cite this

    Fujigaki, H., Saito, K., Lin, F., Fujigaki, S., Takahashi, K., Martin, B. M., Chen, C. Y., Masuda, J., Kowalak, J., Takikawa, O., Seishima, M., & Markey, S. P. (2006). Nitration and inactivation of IDO by peroxynitrite. Journal of Immunology, 176(1), 372-379. https://doi.org/10.4049/jimmunol.176.1.372