Regulation by mitochondrial superoxide and NADPH oxidase of cellular formation of nitrated cyclic GMP: Potential implications for ROS signalling

Khandaker Ahtesham Ahmed, Tomohiro Sawa, Hideshi Ihara, Shingo Kasamatsu, Jun Yoshitake, Md Mizanur Rahaman, Tatsuya Okamoto, Shigemoto Fujii, Takaaki Akaike

Research output: Contribution to journalArticlepeer-review

52 Citations (Scopus)


8-Nitro-cGMP (8-nitroguanosine 3′,5′-cyclic monophosphate) is a nitrated derivative of cGMP, which can function as a unique electrophilic second messenger involved in regulation of an antioxidant adaptive response in cells. In the present study, we investigated chemical and biochemical regulatory mechanisms involved in 8-nitro-cGMP formation, with particular focus on the roles of ROS (reactive oxygen species). Chemical analyses demonstrated that peroxynitrite-dependent oxidation and myeloperoxidase-dependent oxidation of nitrite in the presence of H 2O 2 were two major pathways for guanine nucleotide nitration. Among the guanine nucleotides examined, GTP was the most sensitive to peroxynitrite-mediated nitration. Immunocytochemical and tandem mass spectrometric analyses revealed that formation of 8-nitro-cGMP in rat C6 glioma cells stimulated with lipopolysaccharide plus pro-inflammatory cytokines depended on production of both superoxide and H 2O 2. Using the mitochondria-targeted chemical probe MitoSOX™ Red, we found that mitochondria-derived superoxide can act as a direct determinant of 8-nitro-cGMP formation. Furthermore, we demonstrated that Nox2 (NADPH oxidase 2)-generated H 2O 2regulated mitochondria-derived superoxide production, which suggests the importance of cross-talk between Nox2-dependent H 2O 2 production and mitochondrial superoxide production. The results of the present study suggest that 8-nitro-cGMP can serve as a unique second messenger thatmay be implicated in regulating ROS signalling in the presence of NO.

Original languageEnglish
Pages (from-to)719-730
Number of pages12
JournalBiochemical Journal
Issue number2
Publication statusPublished - 2012 Jan 15


  • Mitochondrion
  • Nitric oxide
  • Oxidative stress
  • Peroxynitrite
  • Reactive oxygen species (ROS)
  • Reactive oxygen species (ROS) signalling

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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