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. 2001 Mar;12(3):539–549. doi: 10.1091/mbc.12.3.539

Figure 6.

Figure 6

The Srp1p/Cse1p/Gsp1p-GTP complex is sensitive to Yrb1p but resistant to Yrb1p complexed to Gsp1p-GTP. GST-Srp1p (8 μg per reaction) was immobilized on glutathione Sepharose and incubated with 12 μg of Cse1p and 4 μg of Gsp1pQ71L-GTP for 30 min at 4°C. After three washes, bound material was incubated with buffer alone (lane 5), with 5 μg of Yrb1p (lane 6), with a preincubated mixture of Yrb1p and Gsp1pQ71L-GTP (lane 7), or with 10 μg of Gsp1pQ71L-GTP (lane 8). After further incubation for 30 min at 4°C, bound (lanes 5–8) and unbound (lanes 9–12) material was separated by SDS PAGE and analyzed by Coomassie blue staining. The molecular weight markers (M) and protein loads are shown in lanes 1–4.