FIGURE 7:

Coaggregation between Aβ peptides and mitochondrial precursor protein. Precursor protein [³⁵S]Su9(86)-DHFR was incubated for 30 min at 30°C in import buffer in the presence or absence of the indicated amounts of Aβ peptides. After incubation, samples were analyzed by the following techniques, (A) Tricine-SDS–PAGE. Soluble fractions (Supernatant) were separated from the insoluble (Pellet) by centrifugation for 40 min at 123,000 × g at 4°C. Samples were analyzed by tricine-SDS–PAGE. (B) Filter retardation assay. Samples were filtered directly through cellulose acetate and nitrocellulose membranes using a dot blot filtration unit as described in Material and Methods. Proteins bound to both membranes were stained with Ponceau S. Bound Aβ peptides were detected by immunodecoration and the precursor protein by digital autoradiography. (C) BN–PAGE. Samples were loaded on native PAGE as described in Materials and Methods and analyzed by Western blot. The precursor protein signal was detected by digital autoradiography and the Aβ peptides by immunodecoration.