Uptake of oxidized folates by rat liver mitochondria

Folate, dihydrofolate, and methotrexate are rapidly taken up by rat liver mitochondria. The apparent maximal matrix folate concentration is about 2.5-fold that of the suspending medium, whereas dihydrofolate and methotrexate equilibrate across the inner membrane. Fully reduced folates, including tetrahydrofolate, 5-methyltetrahydrofolate, and 5,10-methylenetetrahydrofolate penetrate only the intermembrane space. Addition of dihydrofolate or methotrexate effects a rapid release of pre-loaded folate, and external methotrexate promotes the release of pre-loaded dihydrofolate. The extent of dihydrofolate uptake is enhanced by addition of folate. These results suggest that oxidized folates are transported to the matrix by a carrier-mediated mechanism.