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. 2020 Dec 4;9:e59055. doi: 10.7554/eLife.59055

Figure 3. Ablation of RCK domain reduces the temperature-dependence of MthK.

(A) Representative single-channel recordings of purified MthK PO reconstituted in soybean polar lipids with 1 mM calcium at 20°C (blue) and 36°C (red). Inset shows the enlarged traces. nPo values are shown below each trace; the errors were calculated as SEM from bootstrapping the whole recordings. (B) Open-dwell time histograms at the two different temperatures. (C) Plot of open-dwell times (solid black) and unitary conductance (dashed blue) of MthK PO with respect to temperature. Error bars represents SEM calculated from n = 5 independent patches.

Figure 3—source data 1. Source data for Figure 3B and C.

Figure 3.

Figure 3—figure supplement 1. Purification and single-channel recordings of MthK PO.

Figure 3—figure supplement 1.

(A) SEC profile of MthK IR (black) and MthK PO (red) obtained using Superdex 200 column. Black and red arrowheads mark the peaks for MthK IR and MthK PO respectively. (B) SDS-PAGE analysis of MthK IR before and after trypsin digestion is shown on the left, whereas the peak fraction from SEC of MthK PO is on the right. (C) Extended single-channel recordings of MthK PO at 1 mM calcium. The two arrows mark the region shown in Figure 3A.
Figure 3—figure supplement 1—source data 1. Source data for gel filtration profile comparision of MthK IR and MthK Pore.