Transport of pyridoxine and pyridoxal 5'-phosphate in isolated rat liver mitochondria.
The transport of [14C]pyridoxal-P and [14C]pyridoxine into isolated rat mitochondria was studied by centrifugal filtration. The incubation medium contained 20 mM 2-oxoglutarate and 10 mM inorganic phosphate to inhibit metabolism of pyridoxal-P by the mitochondria. The ratio of [14C]pyridoxine space to [3H]H2O space rapidly attained unity independent of the [14C]pyridoxine concentration in the medium and remained unchanged for up to 90 min of incubation. These data suggest simple passive diffusion for the transport of pyridoxine into the mitochondria. By contrast, the ratio of [14C]pyridoxal-P space to [3H]H2O space rose rapidly to exceed 1 in the first 15 min and continued to rise at a slower rate for as long as it was measured. The accumulation of [14C]pyridoxal-P was not decreased by inhibitors and uncouplers of oxidative phosphorylation. Fractionation of the mitochondria with digitonin revealed that 19 and 340 pmol of [14C]pyridoxal-P/mg of protein were taken up by the mitochondria at 15 and 60 min of incubation, respectively. Most of the uptake in the first 15 min occurred in the intermembrane space, whereas the largest increase of [14C]pyridoxal-P between 15 and 60 min of incubation appeared in the matrix fraction. Significant binding of the [14C]pyridoxal-P to proteins in the two compartments was demonstrated by gel filtration. These data indicate that pyridoxal-P can rapidly enter the intermembrane space of isolated mitochondria, but its penetration into the matrix occurs at a slower and more sustained rate (i.e. 9-16 pmol/h/mg of protein). It is concluded that the transport of pyridoxal-P into isolated rat liver mitochondria is energy-independent and is most consistent with passive diffusion facilitated by protein binding once this coenzyme enters the different compartments of the mitochondria.
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D011732:Pyridoxal Phosphate / Q000378:metabolism*; D011736:Pyridoxine / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D014475:Uncoupling Agents / Q000494:pharmacology
D014475:Uncoupling Agents; D011732:Pyridoxal Phosphate; D011736:Pyridoxine