The Different Routes of Calcium Efflux from Liver Mitochondria

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Abstract
Calcium efflux from liver mitochondria, induced by an uncoupler during incubation at 20 degrees C, is largely inhibited by the prior addition of ruthenium red or EGTA. The inhibition by EGTA (i.e. by chelation of Ca2+) is impaired by the presence of EDTA (i.e. by chelation of Mg2+), and it is completely abolished by addition of spermine. In contrast, the inhibition of calcium efflux at 20 degrees C by ruthenium red is unaffected by EGTA or spermine. This latter pathway of calcium efflux therefore represents the reversal of the calcium uniporter. During incubation at 30 degrees C, uncoupler-induced calcium efflux is incompletely inhibited by ruthenium red, and the residual calcium efflux occurs via membrane transition. The kinetics of this process as well as its exceptionally strong dependence on temperature constitute the main evidence for considering that membrane transition modifies the uniporter, and that the modified uniporter is responsible for the residual calcium efflux. It was shown that the route of ruthenium red-insensitive calcium efflux from energized mitochondria under standard conditions is the same, irrespective of whether the uniporter is running or is blocked by ruthenium red. The development of methods for the clear experimental separation of these different routes of calcium efflux under different conditions is still critically important.

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