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

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55 Citations (Scopus)

Abstract

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
Volume320
Issue number3
DOIs
Publication statusPublished - 2004 Jul 30

Keywords

  • 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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  • Cite this

    Nomiyama, T., Igarashi, Y., Taka, H., Mineki, R., Uchida, T., Ogihara, T., Choi, J. B., Uchino, H., Tanaka, Y., Maegawa, H., Kashiwagi, A., Murayama, K., Kawamori, R., & Watada, H. (2004). Reduction of insulin-stimulated glucose uptake by peroxynitrite is concurrent with tyrosine nitration of insulin receptor substrate-1. Biochemical and biophysical research communications, 320(3), 639-647. https://doi.org/10.1016/j.bbrc.2004.06.019