Differential sensitivity of guanylyl cyclase and mitochondrial respiration to nitric oxide measured using clamped concentrations.
Nitric oxide (NO) signal transduction may involve at least two targets: the guanylyl cyclase-coupled NO receptor (NO(GC)R), which catalyzes cGMP formation, and cytochrome c oxidase, which is responsible for mitochondrial O(2) consumption and which is inhibited by NO in competition with O(2). Current evidence indicates that the two targets may be similarly sensitive to NO, but quantitative comparison has been difficult because of an inability to administer NO in known, constant concentrations. We addressed this deficiency and found that purified NO(GC)R was about 100-fold more sensitive to NO than reported previously, 50% of maximal activity requiring only 4 nm NO. Conversely, at physiological O(2) concentrations (20-30 microM), mitochondrial respiration was 2-10-fold less sensitive to NO than estimated beforehand. The two concentration-response curves showed minimal overlap. Accordingly, an NO concentration maximally active on the NO(GC)R (20 nm) inhibited respiration only when the O(2) concentration was pathologically low (50% inhibition at 5 microM O(2)). Studies on brain slices under conditions of maximal stimulation of endogenous NO synthesis suggested that the local NO concentration did not rise above 4 nm. It is concluded that under physiological conditions, at least in brain, NO is constrained to target the NO(GC)R without inhibiting mitochondrial respiration.
Bellamy
Tomas C
TC
Wolfson Institute for Biomedical Research, University College London, Gower Street, London, WC1E 6BT, United Kingdom.
Griffiths
Charmaine
C
Garthwaite
John
J
D000818:Animals; D002531:Cerebellum / Q000187:drug effects / Q000502:physiology*; D004048:Diethylamines / Q000494:pharmacology; D006162:Guanylate Cyclase / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008928:Mitochondria / Q000502:physiology*; D009569:Nitric Oxide / Q000494:pharmacology / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009589:Nitrogen Oxides; D010101:Oxygen Consumption / Q000187:drug effects*; D018408:Patch-Clamp Techniques; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology*; D000071756:Soluble Guanylyl Cyclase; D014366:Tryptophan Oxygenase / Q000037:antagonists & inhibitors
D004048:Diethylamines; D020030:Nitric Oxide Donors; D009589:Nitrogen Oxides; D018160:Receptors, Cytoplasmic and Nuclear; D009569:Nitric Oxide; C087229:diethylamine dinitric oxide adduct; D014366:Tryptophan Oxygenase; D006162:Guanylate Cyclase; D000071756:Soluble Guanylyl Cyclase