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

Abstract 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 aminotransferase 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.