Fig. 4.

Mapping the part of GCC88 that can capture vesicles. a Schematic diagram of human GCC88 along with plots for the predicted degree of coiled-coil and disorder along its length. Also shown is the mitochondrial form as in Fig. 1a. b Summary of the vesicle capture activity of the indicated truncations and chimeras of mitochondrial GCC88. Capture at mitochondria was assayed by immunofluorescent staining of the integral membrane proteins CD-MPR, CI-MPR and Vti1a. A plus sign indicates that capture of all three markers was similar to the wild-type protein, a minus sign indicates that no significant capture was observed. c Alignment of the N-terminus of human GCC88 with that from the indicated species. Bird, G. gallus; frog, X. tropicalis; fish D. rerio; urchin, S. purpuratus; fly, D. melanogaster; oyster, C. gigas; hydra, H. vulgaris; anemone, N. vectinis. d, e Confocal micrographs of HeLa cells expressing the indicated GCC88 variants and stained for the hemagglutinin tag on the GCC88 chimera as well as for both CD-MPR (in vesicles captured by GCC88) and ZFPL1 (a cis-Golgi protein that is not captured). Key constructs from the set shown in (b) are included, with similar results also obtained using the vesicle markers CI-MPR and Vti1a. Scale bars 10 μm