Fig. 6
Litreol binds to an inner mitochondrial membrane protein from bovine heart mitochondria with a molecular weight of approximately 27 kD. Mitochondria isolated from the bovine heart were treated with or without 5 μM 3H-litreol and then with cold litreol, according to the “Methods” section. After submitochondrial fractionation, the proteins of the internal membranes (IM) and mitoplasts (MP) were analyzed by SDS-PAGE. The fractions from mitochondria treated with 3H-litreol are indicated. Two identical gels were generated: one was stained with Coomassie blue (left panel) and then subjected to fluorography (middle panel). The proteins in the other gel were transferred to a nitrocellulose membrane and developed by Western blotting using an anti-cytochrome c1 antiserum (right panel). A radioactive band (arrow) was observed in the mitoplast and inner membrane fractions and showed electrophoretic mobility similar to that of cytochrome c1 (27 kD). St: prestained marker