Translocation of proteins into rat liver mitochondria

<jats:p>The precursor polypeptides of a large subunit of succinate dehydrogenase and ornithine aminotransferase (the enzymes which are located in the mitochondrial inner membrane and matrix respectively) were synthesized as a larger molecular mass than their mature subunits, when rat liver RNA was translated <jats:italic>in vitro</jats:italic>. These precursor polypeptides were also detected <jats:italic>in vivo</jats:italic> in ascites hepatoma cells (AH‐130 cells). When the <jats:sup>35</jats:sup>S‐labeled precursor polypeptides were incubated with isolated rat liver mitochondria at 30°C in the presence of an energy‐generating system, these two precursors were converted to their mature size and the <jats:sup>35</jats:sup>S‐labeled mature‐size polypeptides associated with mitochondria. Furthermore, these mature‐size polypeptides were recovered from their own locations, the inner mitochondrial membrane and the matrix.</jats:p><jats:p>The precursor of ornithine aminotransferase incubated with rat liver mitochondria at 0°C was specifically and tightly bound to the surface of the mitochondria even in the presence of an uncoupler of oxidative phosphorylation. This precursor, bound to the mitochondria, was imported into the matrix when the mitochondria were reisolated and incubated at 30°C in the presence of an energy‐generating system, suggesting that a specific receptor may be involved in the binding of the precursor.</jats:p><jats:p>The processing enzyme for both precursor polypeptides seemed to be located in the mitochondrial matrix and was partially purified from the mitochondria. A metal‐chelating agent strongly inhibited the processing enzyme and the inhibition was recovered by the addition of Mn<jats:sup>2+</jats:sup> or Co<jats:sup>2+</jats:sup>.</jats:p>