Figure 8

C-Raf blocks VDAC reconstitution into membranes. Membrane conductance as a function of time after the addition of 50 ng/ml VDAC (full squares) and 50 ng/ml VDAC pre-incubated with 150 ng/ml GST-C-RafDD (full points) to DiphPC membranes (left-hand side arrow). The aqueous phase contained 300 mM KCl supplemented with kinase buffer. 15 minutes after addition of VDAC, when the conductance was stationary 150 ng/ml GST-C-RafDD was added to both sides of the membrane (full squares, right-hand side arrow). The temperature was 20°C and the applied voltage was 20 mV. Note that the membrane conductance increase was inhibited when VDAC was preincubated with GST-C-RafDD (full squares), while this protein had no influence on already existing channels (full points).