Spegazzinine, a new inhibitor of mitochondrial oxidative phosphorylation

The effects of spegazzinine, a dihydroindole alkaloid, on mitochondrial oxidative phosphorylation were studied. Spegazzinine inhibited coupled respiration and phosphorylation in rat liver mitochondria. The I50 was 120 microM. Uncouplers released the inhibition of coupled respiration. Arsenate-stimulated mitochondrial respiration was partially inhibited by spegazzinine. The stimulation of mitochondrial respiration by Ca2+ and the proton ejection associated with the ATP-dependent Ca2+ uptake were not affected by the alkaloid. Oxidative phosphorylation and the Pi-ATP exchange reaction of phosphorylating beef heart submitochondrial particles were strongly inhibited by spegazzinine (I50, 50 microM) while the ATP-dependent reactions, reduction of NAD+ by succinate and the pyridine nucleotides transhydrogenase were less sensitive (I50, 125 microM). Oxygen uptake by submitochondrial particles was not affected. The 2,4-dinitrophenol-stimulated ATPase activity of rat liver mitochondria was not affected by 300 microM spegazzinine, a concentration of alkaloid that completely inhibited phosphorylation. However, higher concentrations of spegazzinine did partially inhibit it. The ATPase activities of submitochondrial particles, insoluble and soluble ATPases were also partially inhibited by high concentrations of spegazzinine. The inhibitory properties of spegazzinine on energy transfer reactions are compared with those of oligomycin, aurovertin and dicyclohexylcarbodiimide. It is concluded that spegazzinine effects are very similar to the effects of aurovertin and that its site of action may be the same or near the site of aurovertin.