Extended Data Fig. 8: Structural characterization of the OSH6 interaction with lipid nanodiscs.
From: Structural basis for lipid transport at membrane contact sites by the IST2–OSH6 complex
a, Representative CTF-corrected micrograph for OSH6ND (n = 19.283). b, 2D classes for OSH6ND calculated in cryoSPARC representing the OSH6-bound fraction of nanodiscs used for initial 3D reconstruction. c, Data processing workflow for OSH6ND performed in cryoSPARC and Relion v5. d, Results of 2D classification without alignment of refined particles in the final map performed in Relion v5. e, GSFSC curve of final map displayed in (c) computed in Relion v5. f, Euler angle distribution for final 3D reconstruction shown for two orientations of the map. g, Positioning of the OSH6 model into the OSH6ND map. Left, correlation between the experimentally determined map (shown in c) and 100 representative placements covering all orientations of a density map calculated from the OSH6 structure. The initial placement was followed by a local optimization to improve the orientation resulting in 50 unique clusters. The red data point represents the fit shown in Fig. 4d. Center, close-up of the top four hits obtained. Right, histogram of hits within the top four unique clusters showing the largest population for the top hit. h, i, Size-exclusion chromatography analysis of OSH6 and MSP2N2 in the absence of lipids. h, Data points show normalized band intensity from SDS-PAGE analysis of SEC fractions shown in i, collected at the indicated retention volumes. h,i, The experiment was performed once.
