Peroxynitrite formation from macrophage-derived nitric oxide

Arch Biochem Biophys. 1992 Nov 1;298(2):446-51. doi: 10.1016/0003-9861(92)90433-w.


Peroxynitrite formation by rat alveolar macrophages activated with phorbol 12-myristate 13-acetate was assayed by the Cu,Zn superoxide dismutase-catalyzed nitration of 4-hydroxyphenylacetate. The inhibitor of nitric oxide synthesis N-methyl-L-arginine prevented the Cu,Zn superoxide dismutase-catalyzed nitration of 4-hydroxyphenylacetate by stimulated macrophages, while Cu-depleted Zn superoxide dismutase did not catalyze the formation of 3-nitro-4-hydroxyphenylacetate either in vitro or in the presence of activated macrophages. The rate of phenolic nitration by activated macrophages was 9 +/- 2 pmol x 10(6) cells-1 x min-1 (mean +/- STD). Only 8% of synthetic peroxynitrite was trapped by superoxide dismutase, which suggested that the rate of peroxynitrite formation may have been as high as 0.11 nmol x 10(6) cells-1 x min-1. This upper estimate was consistent with N-methyl-L-arginine increasing the amount of superoxide detected with cytochrome c by 0.12 nmol x 10(6) cells-1 x min-1. The rate of nitrite and nitrate accumulation was 0.10 +/- 0.001 nmol x 10(6) cells-1 x min-1, suggesting that the majority of nitric oxide produced by activated macrophages may have been converted to peroxynitrite. The formation of a relatively long lived, strong oxidant from the reaction of nitric oxide and superoxide in activated macrophages may contribute to inflammatory cell-mediated tissue injury.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cattle
  • Erythrocytes / enzymology
  • Kinetics
  • Macrophages, Alveolar / drug effects
  • Macrophages, Alveolar / metabolism*
  • Nitrates / metabolism*
  • Nitric Oxide / metabolism*
  • Oxygen Consumption
  • Rats
  • Rats, Sprague-Dawley
  • Superoxide Dismutase / blood
  • Superoxide Dismutase / metabolism*
  • Superoxides / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology


  • Nitrates
  • Superoxides
  • peroxynitric acid
  • Nitric Oxide
  • Superoxide Dismutase
  • Tetradecanoylphorbol Acetate