Processing-independent in vitro translocation of cytochrome P-450(SCC) precursor across mitochondrial membranes.
In the presence of a membrane-permeable metal chelator, bovine adrenal cortex mitochondria imported P-450(SCC) precursor without processing of the amino-terminal extension peptide. The imported precursor was bound to the matrix side surface of the inner membrane. When the inhibition due to the metal chelator was removed, the imported precursor was processed to the mature form. Unprocessed precursor was also detected in mitochondria when the import reaction was carried out at relatively low temperature. These results suggest that the translocation of P-450(SCC) precursor across mitochondrial membranes is independent of its processing to the mature form. Both membrane-bound and solubilized P-450(SCC) could be cleaved by trypsin into two fragments with molecular weights of 29 kDa and 26 kDa, respectively, suggesting a two-domain structure of the molecule. The in vitro-imported and processed P-450(SCC) was also cleaved by trypsin in the same way. This finding indicated that the in vitro-imported and processed P-450(SCC) has the same conformation as the native form.
D000302:Adrenal Cortex / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D002417:Cattle; D002614:Chelating Agents / Q000494:pharmacology; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism*; D007425:Intracellular Membranes / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D008970:Molecular Weight; D010088:Oxidoreductases / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D010618:Phenanthrolines / Q000494:pharmacology; D011487:Protein Conformation; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D014357:Trypsin / Q000378:metabolism
D002614:Chelating Agents; D010446:Peptide Fragments; D010618:Phenanthrolines; D011498:Protein Precursors; D010088:Oxidoreductases; D002786:Cholesterol Side-Chain Cleavage Enzyme; D014357:Trypsin; C025205:1,10-phenanthroline