Figure 1.
Mcl-1 binds strongly to VDAC1 and 3 and binds to VDAC1 with greater apparent affinity than Bcl-xL. (a) Human Mcl-1 expressed in MEF cells and detected by western blot after pulldown by GST-fusion proteins of VDAC1, 2 or 3 is shown in the upper lanes with loading control blots of GST depicted below. (b) GST-fusion proteins of VDAC1, 2 or 3 detected by anti-GST antibody after pulldown by purified His-tagged Mcl-1 is shown in the upper lanes with the loading control blots of Mcl-1 depicted below. Lanes labeled BSA are negative controls showing GST-VDACs are not pulled down by beads alone. (c) Western blot detection of Mcl-1 and Bcl-xL pulled down by GST-VDAC1 from MEF cell lysates pretreated with control or VDAC-based peptides (N-ter and L14-15). (d) Western blot of recombinant purified Mcl-1 and Bcl-xL (0.2 μg) pulled down by GST-VDAC1. All blots are representative of at least two independent experiments