Figure 10.

The role of the C-terminal tail of Emp46p in recruitment into the prebudding complex. Microsomes prepared from cells expressing either wild-type or mutated Emp46p were incubated with GST-Sar1p and the purified Sec23/24p complex in the presence of GMP-PNP or GDP-βS. Subsequently, digitonin-soluble prebudding complexes were recovered on glutathione beads. Emp46p or Sec22p in the prebudding complex was detected by immunoblotting. T represents 0.5% of the total solubilized membranes. Each construct is denoted as described in the legend to Figure 6B.