Figure 5.

Binding of VCP to Atx-3 influences the assembly of polyQ-containing protein aggregates. (A) VCP stimulates the formation of Atx-3 aggregates in vitro. 3 μM GST-Atx71Q(242–360) or GST-Atx68Q(242–360)²⁸²HNHH was incubated with 1 μM VCP or 1 μM ovalbumin in the presence of PP. The assembly of SDS-stable Atx-3 protein aggregates was monitored by filtration using an anti-Atx (CT1) antibody. The sample with the highest signal intensity was arbitrarily set as 100%. (B) Effect of different VCP concentrations on Atx-3 aggregation in vitro. GST fusion proteins were incubated for 6 h with PP and different amounts of VCP. The assembly of SDS-insoluble Atx71Q(242–360) and Atx68Q(242–360)²⁸²HNHH aggregates was monitored by filtration using the CT1 antibody. Relative amounts of aggregates were quantified by phosphorimager densitometry. The X-axis of the diagram shows the ratio of VCP to Atx-3.