Production of nitric oxide by mitochondria

Cecilia R Giulivi, Juan José Poderoso, Alberto Boveris

Research output: Contribution to journalArticle

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Abstract

The production of NO· by mitochondria was investigated by electron paramagnetic resonance using the spin-trapping technique, and by the oxidation of oxymyoglobin. Percoll-purified rat liver mitochondria exhibited a negligible contamination with other subcellular fractions (1-4%) and high degree of functionality (respiratory control ratio = 5-6). Toluene- permeabilized mitochondria, mitochondrial homogenates, and a crude preparation of nitric oxide synthase (NOS) incubated with the spin trap N- methyl-D-glucamine-dithiocarbamate-Fe(II) produced a signal ascribed to the NO· spin adduct (g = 2.04; a(N) = 12.5 G). The intensity of the signal increased with time, protein concentration, and L-Arg, and decreased with the addition of the NOS inhibitor N(G)-monomethyl-L-arginine. Intact mitochondria, mitochondrial homogenates, and submitochondrial particles produced NO· (followed by the oxidation of oxymyoglobin) at rates of 1.4, 4.9, and 7.1 nmol NO· x (min·mg protein)-1, respectively, with a K(m) for L-Arg of 5-7 μM. Comparison of the rates of NO· production obtained with homogenates and submitochondrial particles indicated that most of the enzymatic activity was localized in the mitochondrial inner membrane. This study demonstrates that mitochondria are a source of NO·, the production of which may effect energy metabolism, O2 consumption, and O2 free radical formation.

Original languageEnglish (US)
Pages (from-to)11038-11043
Number of pages6
JournalJournal of Biological Chemistry
Volume273
Issue number18
DOIs
StatePublished - May 1 1998
Externally publishedYes

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Mitochondria
Nitric Oxide
Submitochondrial Particles
Nitric Oxide Synthase
Spin Trapping
Subcellular Fractions
Liver Mitochondrion
Electron Spin Resonance Spectroscopy
Toluene
Mitochondrial Membranes
Oxidation
Energy Metabolism
Free Radicals
Arginine
Proteins
Liver
Paramagnetic resonance
Rats
Contamination
Membranes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Production of nitric oxide by mitochondria. / Giulivi, Cecilia R; Poderoso, Juan José; Boveris, Alberto.

In: Journal of Biological Chemistry, Vol. 273, No. 18, 01.05.1998, p. 11038-11043.

Research output: Contribution to journalArticle

Giulivi, Cecilia R ; Poderoso, Juan José ; Boveris, Alberto. / Production of nitric oxide by mitochondria. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 18. pp. 11038-11043.
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