Fig. 5.

TRC35 binding precludes karyopherin α binding to Bag6. Recombinantly purified Bag6C131–6xHis⋅Ubl4A (500 pmol) was first incubated with either GST⋅TRC35 or MBP⋅KPNA2 (500 pmol). The resulting GST⋅TRC35–Bag6–Ubl4A complex was incubated with glutathione resin beads, and increasing amounts of MBP⋅KPNA2 (50, 250, or 1,000 pmol) was added. The ability of MBP⋅KPNA2 to displace Bag6–Ubl4A from GST⋅TRC35 was examined by eluting the GST⋅TRC35 and bound Bag6–Ubl4A from the glutathione resin. The opposite experiment was also carried out using amylose resin beads.