Fig. 4.

The bilayer-facing Glu is a critical determinant of pacCer photolabeling of VDACs. a Human VDAC1, VDAC1^E73Q, VDAC2, and VDAC2^E84Q were produced in E. coli, purified, reconstituted in liposomes, and incubated for 30 min at 37 °C with liposomes containing 1 mol% of pacCer or photoactive and clickable analogs of diacylglycerol (pacDAG), phosphatidylcholine (pacPC), phosphatidylethanolamine (pacPE), or cholesterol (pacChol). Samples were UV irradiated and then click-reacted with AF647-N₃, subjected to SDS-PAGE, and analyzed by IGF and CB staining. b Quantitative analysis of relative labeling efficiencies of reconstituted VDAC1, VDAC1^E73Q, VDAC2, and VDAC2^E84Q with pacLipids as indicated in a. Data are means ± s.d.; n ≥ 3. Source data