Electrophysiological and cryo-EM analysis.
A, ΔCR_PrP with a membrane anchor induces spontaneous inward currents in HEK293 cells. Representative whole-cell patch clamp recordings are shown. HEK293 cells were clamped from a holding potential of 0 mV to a test potential of −100 mV for 10 s every 30 s. After a control phase, the indicated proteins were added directly to the bath solution. In the presence of FL_PrP-MA (left) and ΔCR_PrP without a membrane anchor (middle), no or only weak pore forming activity was observed, whereas ΔCR_PrP-MA induced large sodium (top right) and calcium (bottom right) inward currents. B, cryo-EM images of POPG vesicles (arrows) mixed with ΔCR_PrP-MA (left) and POPG vesicles alone (right). The scale bar is 100 nm.