Purification and identification of a cytosolic factor required for import of precursors of mitochondrial proteins into mitochondria.

A cytosolic factor required for import of the precursor of mitochondrial protein into mitochondria was purified to homogeneity from a rabbit reticulocyte lysate by affinity column chromatography using a synthetic peptide containing the presequence of ornithine amino-transferase as a ligand. The molecular mass of the purified protein was estimated as 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The import of precursors of ornithine aminotransferase and sulfite oxidase into mitochondria was inhibited by anti-28-kDa protein IgG raised in guinea pigs. This antibody also blocked the binding of these precursors to mitochondria. These results suggest that the 28-kDa protein is an essential component of the import machinery in the cytosol and that anti-28-kDa protein IgG blocked the binding of the precursor of ornithine aminotransferase to mitochondria, but not the penetration step. Therefore, the 28-kDa protein may be a factor that should be named the "targeting factor" for import of mitochondrial protein.