In vitro degradation of mitochondrial proteins by ATP-dependent protease in bovine adrenal cortex.
We have examined in vitro degradation of mitochondrial proteins by ATP-dependent protease in bovine adrenal cortex. Purified ATP-dependent protease degraded at least five proteins in vitro in the presence of ATP and Triton X-100 using mitochondrial fraction as a substrate. Two of the degraded proteins were identified as P-450scc and adrenodoxin reductase by immunoblotting. No degradation of P-45011 beta or adrenodoxin was detected. Purified P-450scc and adrenodoxin reductase were also degraded. By analyzing degradation products of P-450scc we identified 49 peptides and 59 cleavage sites. The size of the products was between 3 and 18 amino acid residues. The protease preferentially cleaved the carboxy-side of Leu, Phe, Ala, Val, Met, and some other amino acids. Since the protease (a matrix enzyme) is accessible to P-450scc and adrenodoxin reductase localized on the matrix side of inner membrane, the present results suggest that the protease might participate in the turnover of these two proteins and some other mitochondrial proteins.
Watabe
S
S
Radioisotope Research Institute, St. Marianna University School of Medicine, Kawasaki.
Hara
T
T
Kohno
H
H
Hiroi
T
T
Yago
N
N
Nakazawa
T
T
D049069:ATP-Dependent Proteases; D000302:Adrenal Cortex / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000737:chemistry / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity
D006360:Heat-Shock Proteins; D010446:Peptide Fragments; D011506:Proteins; D002786:Cholesterol Side-Chain Cleavage Enzyme; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases