Journal: Microbial Cell Factories

Article Title: Purification and initial characterization of Plasmodium falciparum K + channels, PfKch1 and PfKch2 produced in Saccharomyces cerevisiae

doi: 10.1186/s12934-020-01437-7

Figure Lengend Snippet: Functional reconstitution of purified PfKch1 1−1094 -GFP and GFP-PfKch2 fl into lipid bilayers. Solubilized and purified PfKch1 1−1094 -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 PfKch1 1−1094 -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.

Article Snippet: Planar lipid bilayers were generated from the GUVs on borosilicate glass chips with a resistance of 2–5 MΩ using the Nanion Port-a-Patch system (see Materials and Methods).

Techniques: Functional Assay, Purification