ATP-dependent spectral response of oxonol VI in an ATP-Pi exchange complex.
Energy transduction in an ATPase complex (complex V) has been studied in two reactions catalyzed by this system, i.e., ATP-dependent spectral shift of oxonol VI, and ATP-Pi exchange activity. Aurovertin alone inhibits 50% of the oxonol shift at 2 microM, and no further inhibition occurs at up to 12 microM. In combination with even weakly effective uncouplers, 4 microM aurovertin fully abolishes the oxonol response. No such effects are observed in the presence of oligomycin and uncouplers. No pH gradient is detectable by quenching of 9-amino-6-chloro-2-methoxyacridine; and nigericin is without effect on the oxonol response. Valinomycin is inhibitory even in the absence of added potassium, due to ammonium ions introduced during the purification steps. Thiocyanate inhibits the dye response by only 10-27%, depending on the preparation. The extent of the oxonol response depends on the ATP/ADP ratio rather than the phosphorylation potential. The dye response in the ATPase complex is 4-7-times less sensitive to bile salts than in submitochondrial particles. The inhibition by cardiolipin can be reversed by the addition of phospholipids. The possibility is discussed that the oxonol response in the ATPase complex reflects, at least in part, a more local, ATP-dependent and energy-related process.
D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology; D002474:Cell-Free System; D005456:Fluorescent Dyes; D007476:Ionophores / Q000494:pharmacology; D007555:Isoxazoles; D008928:Mitochondria / Q000502:physiology*; D025261:Mitochondrial Proton-Translocating ATPases; D010080:Oxazoles; D010743:Phospholipids / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D001647:Bile Acids and Salts; D005456:Fluorescent Dyes; D007476:Ionophores; D007555:Isoxazoles; D010080:Oxazoles; D010743:Phospholipids; D013439:Sulfhydryl Reagents; C021983:complex V (mitochondrial oxidative phosphorylation system); C031184:oxonol VI; D025261:Mitochondrial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases