Nitration of the mitochondrial complex I subunit NDUFB8 elicits RIP1- and RIP3-mediated necrosis

Christiana W. Davis, Brian J. Hawkins, Subbiah Ramasamy, Krishna M. Irrinki, Bruce A. Cameron, Khalid Islam, Varsha P. Daswani, Patrick J. Doonan, Yefim Manevich, Muniswamy Madesh

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75 Scopus citations


Nitric oxide (NO) and other reactive nitrogen species target multiple sites in the mitochondria to influence cellular bioenergetics and survival. Kinetic imaging studies revealed that NO from either activated macrophages or donor compounds rapidly diffuses to the mitochondria, causing a dose-dependent progressive increase in NO-dependent DAF fluorescence, which corresponded to mitochondrial membrane potential loss and initiated alterations in cellular bioenergetics that ultimately led to necrotic cell death. Cellular dysfunction is mediated by an elevated 3-nitrotyrosine signature of the mitochondrial complex I subunit NDUFB8, which is vital for normal mitochondrial function as evidenced by selective knockdown via siRNA. Overexpression of mitochondrial superoxide dismutase substantially decreased NDUFB8 nitration and restored mitochondrial homeostasis. Further, treatment of cells with either necrostatin-1 or siRNA knockdown of RIP1 and RIP3 prevented NO-mediated necrosis. This work demonstrates that the interaction between NO and mitochondrially derived superoxide alters mitochondrial bioenergetics and cell function, thus providing a molecular mechanism for reactive oxygen and nitrogen species-mediated alterations in mitochondrial homeostasis.

Original languageEnglish (US)
Pages (from-to)306-317
Number of pages12
JournalFree Radical Biology and Medicine
Issue number2
StatePublished - Jan 15 2010



  • Bioenergetics
  • Complex I
  • Endothelium
  • Mitochondria
  • Necrosis
  • Nitric oxide
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Davis, C. W., Hawkins, B. J., Ramasamy, S., Irrinki, K. M., Cameron, B. A., Islam, K., Daswani, V. P., Doonan, P. J., Manevich, Y., & Madesh, M. (2010). Nitration of the mitochondrial complex I subunit NDUFB8 elicits RIP1- and RIP3-mediated necrosis. Free Radical Biology and Medicine, 48(2), 306-317.