1. MOSPD2 is an ER‐resident protein able to tether synthetic vesicles by binding the FFAT motif
MOSPD2 is an ER‐resident protein able to tether synthetic vesicles by binding the FFAT motif HeLa/GFP‐MOSPD2 cells were labeled with anti‐Calnexin antibodies (top; magenta) or with anti‐VAP‐A antibodies (bottom; magenta).Pearson correlation coefficients between GFP‐MOSPD2 and Calnexin (left) or GFP‐MOSPD2 RD/LD and Calnexin (right) staining are shown. Each dot represents a single cell (20 cells from three independent experiments). Means and error bars (SD) are shown. Mann–Whitney test.HeLa cells co‐expressing GFP‐MOSPD2 RD/LD (green) and mCherry‐MOSPD2 (magenta).Description of the liposome aggregation assay experimental strategy. LA liposomes are decorated with an FFAT peptide owing to covalent links with MPB‐PE lipids, and mixed with LB liposomes covered by 6His‐tagged MOSPD2 MSP domain attached to DOGS‐NTA‐Ni2+.Aggregation assays in real time. LA liposomes (50 μM total lipids) decorated with conventional FFAT peptide (380 nM) were mixed with LB liposomes (50 μM total lipids) covered with the wild‐type (top; 760 nM) or the RD/LD mutant (bottom; 760 nM) MSP domain of MOSPD2. Aggregation was followed by dynamic light scattering (DLS). Left panels: mean radius (black dots) and polydispersity (shaded area) over time. Right panels: size distribution before (gray bars) and after (black bars) the reaction. Representative illustration of at least three independent experiments.Data information: In (A and C), the subpanels on the right are higher magnification (3.5×) images of the area outlined in white. The Overlay panel shows merged green and magenta images. The Coloc panel displays a colocalization mask on which pixels where the green and the magenta channels co‐localize are shown in white. Right: Linescan analyses with fluorescence intensities of the green and magenta channels along the white arrow shown on the subpanel Overlay. Scale bars: 10 μm.
Thomas Di Mattia et al. EMBO Rep. 2018;19:e45453
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