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Subcellular fractionation of rat liver cells revealed that a mixture of 14C- and 3H-labelled folic acid was distributed approximately equally between the mitochondria and cytosol 2, 24, 48 and 72 h after oral administration. Subfractionation of liver mitochondria 48 h after oral administration showed that the radioactivity was mainly associated with the inner membrane (27.7%) and matrix (51.5%). Hot-ascorbate extraction of the cell cytosol, mitochondrial inner membrane and matrix showed the majority of folates were present as polyglutamates. Acid treatment of isolated folates from cytosol, inner membrane and matrix produced breakdown products consistent with scission of tetrahydrofolates. The folates isolated in the mitochondrial matrix were bound to protein that had an estimated mol. wt. of 90,000.

The distribution and chemical nature of radioactive folates in rat liver cells and rat liver mitochondria