Generation of superoxide by purified brain nitric oxide synthase

J Biol Chem. 1992 Dec 5;267(34):24173-6.

Abstract

Brain nitric oxide synthase (NOS), which utilizes NADPH and calcium/calmodulin as cofactors for metabolizing L-arginine to nitric oxide (NO) and L-citrulline, contains recognition sites for the flavins FAD and FMN. Using a spin-trapping technique combined with electron spin resonance spectroscopy, we report that brain NOS generates superoxide O2-. in a calcium/calmodulin-dependent manner. The "specific inhibitors" of NOS, NG-monomethyl L-arginine (L-NMMA), and NG-nitro-L-arginine methyl ester (L-NAME), have different effects on O2-. generation. For L-NMMA, O2-. production is unaffected, while for L-NAME, inhibition of this free radical is concentration-dependent.

Publication types

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

MeSH terms

  • Amino Acid Oxidoreductases / genetics
  • Amino Acid Oxidoreductases / isolation & purification
  • Amino Acid Oxidoreductases / metabolism*
  • Animals
  • Arginine / analogs & derivatives
  • Arginine / pharmacology
  • Brain / enzymology*
  • Cell Line
  • Chromatography, Affinity
  • Electron Spin Resonance Spectroscopy / methods
  • Kidney
  • Kinetics
  • NG-Nitroarginine Methyl Ester
  • Nitric Oxide Synthase
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Superoxides / metabolism*
  • Transfection
  • omega-N-Methylarginine

Substances

  • Recombinant Proteins
  • Superoxides
  • omega-N-Methylarginine
  • Arginine
  • Nitric Oxide Synthase
  • Amino Acid Oxidoreductases
  • NG-Nitroarginine Methyl Ester