Fig. 10

Functional reconstitution of purified PfKch1¹⁻¹⁰⁹⁴-GFP and GFP-PfKch2^fl into lipid bilayers. Solubilized and purified PfKch1¹⁻¹⁰⁹⁴-GFP and GFP-PfKch2^fl proteins were reconstituted into giant unilamellar vesicles (GUVs) consisting of 10 mM DPhPC and 1 mM cholesterol and measured in a planar lipid bilayer setup in symmetrical KCl solutions (135 mM) using the Nanion Port-a-Patch system. a The left panel shows the result of incorporation of a single PfKch1¹⁻¹⁰⁹⁴-GFP channel into the bilayer. Currents are recorded at −40 or +40 mV. Closed states of the channel (zero current) are indicated with a dashed line (c) and openings (o) are seen as upward and downward deflections at negative and positive potentials respectively. Sub-conductance states are indicated by red arrows. The right panel shows an IV-plot of single channel events recorded at different voltages ranging from −160 mV to +160 mV. The data are obtained from 7 independent experiments with one or more incorporated channels. b: Single channel currents of GFP-PfKch2^fl were recorded at -40 and +40 mV as described above. The left panel shows the results of incorporation of a single channel and open and closed states are indicated as in a. The right panel shows the IV-plot of single channel events recorded in 6 independent experiments.