title: Cell-free synthesis of mitochondrial ATPase inhibitor precursor and its transport into yeast mitochondria.
abstract: ATPase inhibitor protein, which blocks mitochondrial ATPase activity by forming an enzyme-inhibitor complex, was found to be synthesized as a larger precursor in a cell-free translation system directed by yeast mRNA. Other protein factors, which stabilize latent ATPase by binding to the enzyme-inhibitor complex, were also found to be formed as larger precursors. The precursor of ATPase inhibitor protein was transported into isolated yeast mitochondria and was cleaved to the mature peptide in the mitochondria. Impaired mitochondria lacking phosphorylation activity could not convert the precursor to the mature form. Neither antimycin A nor oligomycin alone exhibited a marked effect on the transport-processing of the precursor by intact mitochondria. However, when antimycin A was added with oligomycin, the transport-processing was markedly inhibited. The processing was also strongly inhibited by an uncoupler, carbonylcyanide p-trifluoro-methoxyphenyl hydrazone. The inhibition by the uncoupler was not relieved by ATP added externally. It is concluded that the transport-processing of precursor proteins requires intact mitochondria with a potential difference across the inner membrane.
mesh_terms: D000818:Animals; D002474:Cell-Free System; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis*; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D000097795:ATPase Inhibitory Protein
chemical_list: D011498:Protein Precursors; D011506:Proteins; D012333:RNA, Messenger; D000097795:ATPase Inhibitory Protein