Fig. 5

Mapping the part of golgin-97 that captures vesicles. a Schematic diagram of human golgin-97 along with plots for the predicted degree of coiled-coil and disorder along its length. Also shown is the mitochondrial form in which the GRIP domain has been replaced with a hemagglutinin (HA) tag and the human monoamine oxidase A transmembrane domain. b Summary of the vesicle capture activity of the indicated truncations and chimeras of golgin-97. 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 golgin-97 with that from the indicated species. Bird, G. gallus; frog, X. tropicalis; fish D. rerio; urchin, S. purpuratus; octopus, O. bimaculoides; bee, A. mellifera; fly, D. melanogaster; centipede, S. maritime. d, e Confocal micrographs of HeLa cells expressing the indicated golgin-97 variants and stained for the HA tag on the golgin-97 chimera as well as for CI-MPR or CD-MPR (in vesicles captured by golgin-97) along with ZFPL1 (a cis-Golgi protein that is not captured). Key constructs from the set shown in (b) are included, with similar results obtained using the vesicle markers CD-MPR, CI-MPR or Vti1a. Scale bars 10 μm