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. 2018 Jul 10;8:10406. doi: 10.1038/s41598-018-28751-w

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Mutational studies for the identification of the location of the two different processes found in O-M gating. (a) Sequence of Kcv_NTS as compared to KcsA. Asterisks indicate conserved residues, colons and periods strongly and weakly similar properties, respectively. (b) Structural model of Kcv_NTS constructed with Swissmodel⁷⁹ with KirBac1.1 (PDB 1P7B⁵⁴) as a template and drawn with UCSF Chimera⁸⁰. The positions of mutations used in this study are indicated in red and blue, respectively. (c) Representative current traces of the mutants Kcv_NTS G77S, G77T, and S42T measured in symmetrical 100 mM KCl at −120 mV (top) and +120 mV (bottom). C labels the closed state. (d,e) Amplitude histograms (grey) of Kcv_NTS (d) and Kcv_NTS S42T (e) at −160 mV. The fit by the model in Fig. 1d with extended beta distributions⁴⁴ is given in black or red. (f,g) Voltage dependence of the rate constants (f) k_OM and (g) k_MO of Kcv_NTS wt (black), Kcv_NTS S42T (red), Kcv_NTS G77S (blue) and G77T (blue open symbols). For comparison, the rate constants for Kcv_NTS wt and NTS G77S in (f) and (g) have been taken from a previous publication¹⁴. Numbers of experiments = 3 (6 for Kcv_NTS wt). If the standard deviations are smaller than the data points they are less than 25% scatter. All currents were recorded in symmetrical 100 mM KCl.