Superoxide and peroxynitrite generation from inducible nitric oxide synthase in macrophages

Yong Xia, Jay L. Zweier

Research output: Contribution to journalArticle

Abstract

Superoxide (O2/-·) and nitric oxide (NO) act to kill invading microbes in phagocytes. In macrophages NO is synthesized by inducible nitric oxide synthase (iNOS, NOS 2) from L-arginine (L-Arg) and oxygen; however, O2/-· was thought to be produced mainly by NADPH oxidase. Electron paramagnetic resonance (EPR) spin trapping experiments performed in murine macrophages demonstrate a novel pathway of O2/-· generation. It was observed that depletion of cytosolic L-Arg triggers O2/-· generation from iNOS. This iNOS-mediated O2/-· generation was blocked by the NOS inhibitor N-nitro-L-arginine methyl ester or by L-Arg, but not by the noninhibitory enantiomer N-nitro-D-arginine methyl ester. In L-Arg-depleted macrophages iNOS generates both O2/-· and NO that interact to form the potent oxidant peroxynitrite (ONOO-), which was detected by luminol luminescence and whose formation was blocked by superoxide dismutase, urate, or L-Arg. This iNOS- derived ONOO- resulted in nitrotyrosine formation, and this was inhibited by iNOS blockade. iNOS-mediated O2/-· and ONOO- increased the antibacterial activity of macrophages. Thus, with reduced L-Arg availability iNOS produces O2/-· and ONOO- that modulate macrophage function. Due to the existence of L-Arg depletion in inflammation, iNOS-mediated O2/-· and ONOO- may occur and contribute to cytostatic/cytotoxic actions of macrophages.

Original languageEnglish (US)
Pages (from-to)6954-6958
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number13
DOIs
StatePublished - Jun 24 1997

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Keywords

  • Cytotoxicity
  • Electron paramagnetic resonance
  • L-arginine
  • NADPH oxidase
  • Nitrotyrosine

ASJC Scopus subject areas

  • Genetics
  • General

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