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J. Biol. Chem., Vol. 259, Issue 1, 154-160, 01, 1984
J Austin and JR Aprille
Unidirectional transport (influx and efflux) of adenine nucleotides in rat
liver mitochondria was examined using carboxyatractyloside to inhibit rapid
exchange of matrix and external adenine nucleotides via the adenine
nucleotide translocase. Influx of adenine nucleotides was
concentration-dependent. ATP was the preferred substrate with a Km of 2.67
mM and V of the preferred substrate with a Km of 2.67 mM and V of 8.33
nmol/min/mg of protein. For ADP, the Km was 14.7 mM and V was 10.8
nmol/min/mg of protein. Efflux of adenine nucleotides was also
concentration-dependent, varying directly as a function of the matrix
adenine nucleotide pool size. Any increase in the influx of adenine
nucleotides was coupled to an increase in efflux. However, as the external
ATP concentration was increased, influx was stimulated to a much greater
extent than was efflux. This imbalance suggested that under certain
conditions adenine nucleotide movement might be coupled to the movement of
an alternate anion such as phosphate. Adenine nucleotide efflux increased
as the external phosphate concentration was varied from 0.5 to 4 mM. Also,
increasing the external phosphate concentration caused adenine nucleotide
influx to decrease, suggesting competition. In the absence of external
adenines and phosphate, no efflux occurred. Both adenine nucleotide influx
and efflux were depressed if Mg2+ was omitted. Adenine nucleotide efflux in
the presence of external phosphate was inhibited much less by lack of Mg2+
than was efflux in the presence of external ATP. This evidence supports a
model in which either adenine nucleotides (probably with Mg2+) or phosphate
can move across the mitochondrial membrane on a single carrier. Net adenine
nucleotide movements can occur when adenine nucleotide movement is coupled
to the movement of phosphate in the opposite direction.
Carboxyatractyloside-insensitive influx and efflux of adenine nucleotides in rat liver mitochondria
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