Figure 6.

Protein–protein interactions of Ist1, Did2, Vta1, and Vps60. All proteins were visualized by Coomassie staining. Panels showing positive interactions are boxed in red. B, bound; UN, unbound. (A) Direct interaction between bacterially expressed Ist1 and Did2 was tested by pulling down GST alone, GST-Vps60, or GST-Did2 with Ist1-His₆–conjugated Ni²⁺-NTA beads. (B) Purified Vta1-His₆ protein was pulled down with GST-, GST-Ist1–, GST-Did2–, or GST-Vps60–conjugated glutathione Sepharose beads. Asterisks denote background bands. (C) Binding of Ist1 to Vps4 was tested by pulling down GST or GST-Ist1 with Vps4-His₆–conjugated Ni²⁺-NTA beads.