Valosin-containing protein (VCP/p97) is capable of unfolding polyubiquitinated proteins through its ATPase domains

VCP promotes chymotrypsin digestion of Ub5-DHFR. The ubiquitinated-DHFR (200 nM) mixed with increasing amount of VCP were incubated at 25 °C for 10 min, then added 5 nM chymotrypsin and incubated at 37 °C for 10 min. The proteins were detected by western blotting with anti-DHFR antibody.