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. 2020 Aug 19;6(34):eaba4897. doi: 10.1126/sciadv.aba4897

Fig. 4. Vps24 filaments bind to negatively charged lipids and dissociate into shorter filament fragments.

Fig. 4

(A) Liposome flotation assays of Vps24 with POPC, POPC/PS, and POPC/DOPS lipid mixtures. In the presence of negatively charged lipids, Vps24 is found in the floating top fraction. Micrographs of negatively stained Vps24 of floating top fraction with (B) POPC/PS and (C) POPC/DOPS reveal filament fragments associated to dried liposomes. (D) Bottom fraction shows solubilized oligomer species. (E) Pellet fraction of Vps24 in the POPC/PS sample. (F) Quantification of filament lengths in the presence and absence of POPC/DOPS. When incubated with POPC/DOPS liposomes, Vps24 filaments shorten and associate with liposomes. (G) Depiction of filament disassembly by interaction with PS-containing liposomes. (H) Pelletation (top) and co-pelletation POPC/DOPS liposome assay of Vps24 mutants (left) (Δ1–9, K5D/K6D, and K133D/E134A/K137D) in comparison with wild-type (wt) Vps24 (right). Tested Vps24 mutants retain the ability to bind POPC/DOPS liposomes.