Figure 5.
Residues 28–52 interact with exomer in vitro. (A) In vitro interaction assay to measure ability of GST-tagged Chs3 fragments to interact with purified exomer complex. Exomer was detected by α-6xHis antibody which recognized His-tagged Chs6. GST-tagged constructs were detected by Coomassie. In the “mock treated” samples, the final exomer purification buffer was added in place of exomer. (B) Chs3 fragment pull-down assay as described in (A) comparing the two halves of the Chs3(1–52) segment. (C) Assay as described in (A) to compare interaction of N- and C-termini, but with bound exomer detected by Coomassie staining. (D) Assay as described in (A) to analyze effect of 41LVR43 → AAA mutation on interaction.