The CH of ATL2-1 interacts with protein-free liposomes. (A and B) The per-residue confidence score (pLDDT) and (C) predicted aligned error (PAE) of the AlphaFold structure prediction of ATL2-1 are shown in Fig. 7 C. (D) The CH of ATL2-1, but not that of ATL2-2, interacts with protein-free liposomes. Protein-free liposomes (0.5 mM) were incubated with synthetic peptides (300 μM) derived from the CH of ATL2-1, the CH of ATL2-2, or the mutant CH of ATL2-1 [ATL2-1 (KKE)], which is reported to lack autoinhibitory activity (Crosby et al., 2022), at 4°C for 2 h. The mixtures were then mixed at a 1:1 ratio with 70% (w/v) Histodenz, layered with 25% (w/v) Histodenz, and topped with RB150 buffer containing 1 mM EDTA. After centrifugation at 250,000 × g at 4°C for 1 h, the sample was divided into six fractions. The top and bottom fractions were analyzed for the presence of peptides by SDS-PAGE followed by Coomassie Brilliant Blue staining. Source data are available for this figure: SourceData FS4.