%0 Journal Article
%T Protocol for the production and reconstitution of VDAC1 for functional assays.
%A Dearden GI
%A Ravishankar V
%A Sakata KT
%A Menon AK
%A Bergdoll L
%J STAR Protoc
%V 5
%N 3
%D 2024 Aug 7
%M 39116198
暂无%R 10.1016/j.xpro.2024.103240
%X The voltage-dependent anion channel (VDAC) is an abundant and multifunctional outer mitochondrial membrane protein, playing key roles in neurodegeneration, apoptosis, and mitochondrial membrane biogenesis. Here, we present a protocol to produce and reconstitute high yields of detergent-solubilized VDAC, expressed as inclusion bodies in E. coli. We describe steps for purification by affinity chromatography and refolding in lauryldimethylamine-N-oxide (LDAO). We then detail procedures for reconstituting VDAC into membrane vesicles to assay its channel and phospholipid scramblase activity via fluorescence-based assays. For complete details on the use and execution of this protocol, please refer to Bergdoll et al.,1 Queralt-Martín et al., 2 and Jahn et al.3.