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. 2018 Sep 11;8:13607. doi: 10.1038/s41598-018-31945-x

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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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Voltage dependences of open probability (a) and the closed-to-open transition energy (b) for V23T MscL recorded in DIBs (blue symbols) and in patch-clamp experiments (orange symbols). The patch data (orange) are simulated for the membrane tension at which the DIB data are collected. The fitting of open probability was done using Eq. 2. Based on the asymmetry of voltage response (a,b) we can deduce the orientation of the MscL channel incorporated in DIBs (c,d). In patches excised from the cytoplasmic membrane (c) the channel remains in its native orientation with its cytoplasmic side facing the bath. The directionality of MscL incorporation into DIBs is as presented in panel (d) and achieved through unilateral introduction of proteoliposomes into the ‘cis’ compartment.