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. 2002 Jun;13(6):1906–1915. doi: 10.1091/mbc.02-02-0025

Figure 5.

Figure 5

The N-terminus of granuphilin binds to Rab3. (A) GST or GST-granuphilin 1–300 were immobilized on glutathione-agarose beads and incubated with purified wild-type Rab3A loaded with GDP or with GTPγS. The proteins remaining associated with the affinity columns were visualized by Western blotting using a monoclonal antibody against Rab3A. (B) HIT-T15 cells were transiently transfected with wild-type Rab3A, Rab4, Rab6, or Rab13. Two days later, the cells were homogenized and the cell lysates incubated either with GDP or GTPγS for 15 min at 30°C. They were then loaded onto GST-granuphilin 1–300 affinity columns. The proteins interacting with GST-granuphilin 1–300 were detected by Western blotting using an antibody against the epitope tag.