Fig. 4. Distribution of GRASP2 depends on its hydrophobic N-terminus. (A) Expression of the N-terminal deletion mutants GRASP2
SA-GFP and GRASP2
SA2-GFP was confirmed using GFP-specific antibodies. A single band of
100 kDa resembling the GFP-fusion protein is recognised in all transgenic parasite lines. (B) Fluorescence microscopy on live parasites that express either GRASP2-GFP or the N-terminal deletion mutants GRASP2
SA-GFP and GRASP2
SA2. Whereas GRASP2-GFP is restricted to tightly defined compartments (a), mutation of the N-terminus either by complete deletion of the hydrophobic stretch (b) or partial removal of its proximal part leaving the extreme N-terminus intact (c) abolishes Golgi targeting and results in a cytoplasmic distribution of the fusion protein. All images show the nucleus in blue (DAPI).