Reduction of insulin-stimulated glucose uptake by peroxynitrite is concurrent with tyrosine nitration of insulin receptor substrate-1

Takashi Nomiyama, Yasuhiro Igarashi, Hikari Taka, Reiko Mineki, Toyoyoshi Uchida, Takeshi Ogihara, Jong Bock Choi, Hiroshi Uchino, Yasushi Tanaka, Hiroshi Maegawa, Atsunori Kashiwagi, Kimie Murayama, Ryuzo Kawamori, Hirotaka Watada

Research output: Contribution to journalArticlepeer-review

56 Citations (Scopus)


Inducible nitric oxide synthetase plays an essential role in insulin resistance induced by a high-fat diet. The reaction of nitric oxide with superoxide leads to the formation of peroxynitrite (ONOO-), which can modify several proteins. In this study, we investigated whether peroxynitrite impairs insulin-signalling pathway. Our experiments showed that 3-(4-morpholinyl)sydnonimine hydrochloride (SIN-1), a constitutive producer of peroxynitrite, dose-dependently inhibited insulin-stimulated glucose uptake. While SIN-1 did not affect the insulin receptor protein level and tyrosine phosphorylation, it reduced the insulin receptor substrate-1 (IRS-1) protein level, and IRS-1 associated phosphatidylinositol-3 kinase (PI-3 kinase) activity. Although SIN-1 did not induce Ser307 phosphorylation of IRS-1, tyrosine nitration of IRS-1 was detected in SIN-1-treated-Rat1 fibroblasts expressing human insulin receptors. Mass spectrometry showed that peroxynitrite induced at least four nitrated tyrosine residues in rat IRS-1, including Tyr939, which is critical for association of IRS-1 with the p85 subunit of PI-3 kinase. Our results suggest that peroxynitrite reduces the IRS-1 protein level and decreases phosphorylation of IRS-1 concurrent with nitration of its tyrosine residues.

Original languageEnglish
Pages (from-to)639-647
Number of pages9
JournalBiochemical and biophysical research communications
Issue number3
Publication statusPublished - 2004 Jul 30
Externally publishedYes


  • Insulin receptor substrate-1
  • Insulin resistance
  • Nitric oxide
  • Oxidative stress
  • Peroxynitrite
  • iNOS

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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