Fig. 5
From: Structural insights of human mitofusin-2 into mitochondrial fusion and CMT2A onset
MFN2 interacts with MFN1 via G domain. a Surface conservation analysis reveals that the G interface is a highly conserved area in mitofusins. b Pull-down assays showing the interaction between MFN2IM and GST-tagged MFN1IM in different nucleotide-loading conditions. c Pull-down assays showing that the interaction between MFN2IM and MFN1IM is dependent on the G interface. d Native PAGE result showing the association between MFN2IM and MBP-fused MFN1IM. e SDS-PAGE showing the purified MFN2IM/MBP-MFN1IM hetero-complex. f MFN2IM stimulates GTP turnover of MFN1IM. Overall, 0.25 μM MFN1IM and 2.5 μM MFN1IM MFN2IM mutants were used. g Liposome tethering assay for wild-type MFN2IM/MFN1IM and G interface mutants. Representative images from five independent experiments are shown. Scale bar, 50 μm. h FRET experiment showing the G domain dimerization-dependent conformational change of MFN2IM in different nucleotide-loading conditions. The time point for the addition of corresponding nucleotides is indicated by an arrow for each panel. nt denotes nucleotide. i FRET experiment showing the conformational change of MFN2IM and MFN1IM upon association via the G domains in GTP- and \({\mathrm{GDP}} \bullet {\mathrm{BeF}}_{3}^{-}\) loading conditions. j A model for the trans association between MFN2IM and MFN1IM. Source data are provided as a Source Data file
