Fig. 4: Ligand-gated LGC dynamically controls cargo transport across the lipid bilayer.

a GUV-dye-influx assay scheme. b A time series of corresponding confocal images of GUVs incubated with cholesterol and Cy3-labeled LGC (magenta) and Atto633 dye (green), dropper represents the time point at which Atto633 dye was added. Top—closed lid LGC does not allow Atto633 dye influx into GUV interior. Middle—dynamically opening the LGC lid with key results in the influx of Atto633 dye into GUV. Bottom—separate experiment in which adding Atto633 to dynamically closed LGC does not lead to any dye influx into GUVs, showing dynamic closing is effective. Scale bars: 10 μm. The data were representative of n = 3 independent experiments. c Current trace of a single LGC-C channel inserted into a planar DPhPC membrane. The trace was recorded at a voltage of +80 mV for the first 5 s, after which the potential was is switched to −80 mV. d Current-voltage (IV) plot showing average current of 12 individual insertions ± standard error of the mean of LGC-C at membrane potentials ranging from −100 to +100 mV in 20 mV steps. e Conductance histogram of 12 individual insertions of LGC-C obtained at −20 mV. Inset shows an example trace on the transition from closed-lid pore to open-lid after addition of 15 nM key. f Current trace of a single LGC-N + Chol channel inserted into a planar DPhPC membrane. The trace was recorded at a voltage of +80 mV for the first 5 s, after which the voltage potential was switched to −80 mV. g Current-voltage (IV) trace showing average current of 17 individual insertions ± standard error of mean of LGC-O at membrane potentials ranging from −100 to +100 mV in 20 mV steps. h Conductance histogram of 19 individual LGC-N + Chol channels recorded at −20 mV. Inset shows an example trace on the transition from open-lid pore to closed-lid after addition of 15 nM reverse key. All electrophysiological experiments were conducted in buffers composed of 1 M KCl, 10 mM HEPES pH 7.6. Source data are provided as a Source Data file.