Skip to main content
. 2016 Apr 5;6:23947. doi: 10.1038/srep23947

Figure 4. Head-to-head comparison of four different Champ constructs in which the proton pump and ASIC are separated by increasing primary-sequence distance.

Figure 4

For each construct: a cartoon illustrates the structure of the two-component construct, confocal images demonstrate fluorescence expression in culture and graphs show the relative magnitude of the peak outward current and the current at the end of the light pulse. A more negative current at the end of the light pulse indicates a larger ASIC component. Insets: representative traces of the current responses to a 1 s pulse of 560 nm light for each two-component construct (timing of light pulse indicated by green horizontal line). All electrophysiological recordings were performed in low HEPES (0.1 mM) Tyrode’s solution. (a) eArch3.0-YFP only control (n = 9). (b) Co-transfection of eArch3.0 and ASIC2a: eArch3.0 is labeled with mCherry and ASIC2a is labeled with YFP to allow identification of both components in a single cell (n = 9). (c) Champ1.0: eArch3.0 and ASIC2a are separated during protein translation by the ribosomal skip sequence, p2A (n = 14). (d) Champ2.0: eArch3.0 and ASIC2a are fused by a 41 amino acid linker sequence (n = 17). (e) Champ3.0: eArch3.0 and ASIC2a are fused by a short linker sequence (23 amino acid membrane trafficking signal, TS) (n = 16).