Fig. 1. MIGA2 directly binds to phospholipids.
a Diagram showing the domain structure of Mus musculus MIGA2. From the N-terminus, the MIGA2 has a transmembrane domain (TM), Coiled-coil domain, FFAT motif, and LD targeting domain. b Schematic diagram of the liposome sedimentation assay. The mMIGA2 fragments were incubated with liposomes for 60 min and separated by centrifugation. c Figure showing the results of the liposome-binding assay performed as described in b. Supernatant (Sup) and precipitant fractions (Pellet) were analyzed by 12% SDS-PAGE and Coomassie blue staining (N = 2 independent experiments). d The fluorescence lipid-binding assay. The purified mMIGA2 proteins were incubated with NBD-PE and subjected to 10% Clear Native-PAGE (CN-PAGE) (N = 2 independent experiments). Coomassie staining (left) and fluorescent detection (right). e Diagram showing the in vitro lipid extraction assay. Liposomes containing NBD-PE were incubated with the mMIGA2 for 60 min. The mixtures were separated by centrifugation and precipitated fractions were analyzed by a fluorescence measurement. f Bar graph showing the results of lipid extraction by mMIGA2 using experiments performed as described in e. Lipid extraction activity was compared to that of the control using one-way ANOVA (N = 3 independent experiments; individual data point shown as dots, bars show mean ± SD). ****p < 0.0001 (mMIGA2161–570 and mMIGA2313–570). Source data are provided in a Source data file.