Abstract
Three different types of mitochondrial poisons (oligomycin, antimycin A, and dinitrophenol) strongly inhibited Na(+)-Ca2+ exchange in aortic myocytes. Exchange activity was assayed as 45Ca2+ uptake that depended on inverting the Na+ gradient and was inhibited by 25 microM dimethylbenzamil. Glucose markedly decreased the inhibition of exchange activity by these three poisons. Glucose also prevented rotenone from inhibiting exchange and depleting cellular ATP. In the absence of glucose, rotenone decreased ATP and exchange activity with half-times of 0.8 and 0.9 min, respectively. Almost eliminating cellular ATP with rotenone maximally inhibited exchange by 80%. Repletion of ATP with glucose substantially restored Na(+)-Ca2+ exchange activity. Ca2+ uptake by organelles, subsequent to entry via exchange for Na+, does not appear to contribute significantly to exchange activity as assayed in intact myocytes. The specific activity of Na(+)-Ca2+ exchange was approximately 30 nmol.min-1.mg protein-1. These findings suggest that ATP modulates exchange activity and that there are approximately 150,000 Na(+)-Ca2+ exchangers per cell, assuming that the turnover number is 1,000 s-1.
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