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. 2017 Dec 8;7:17229. doi: 10.1038/s41598-017-17396-w

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© The Author(s) 2017

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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Patch-clamp recording from purified MscL mutant V120C reconstituted into liposomes. (A) The MscL-V120C mutant current recorded upon the application of 10 mmHg suction steps onto the patch area. The channel currents (top) and the negative pressure applied to the inside-out azolectin liposome patch through the patch pipette (bottom) are shown. (B) Activation of the mutant channel by application of 3 μM LPC in the patch pipette, which is much less than 25 mol% used in EPR experiments (Fig. 4). (Note that it took ∼2 min for LPC to diffuse inside the pipette to reach and incorporate in the monounsaturated (18:1) POPC liposome patch and activate the channel.) The channel activity is characterized by brief channel openings. Expanded views show the openings of the MscL-V120C mutant in the presence (left) and absence of suction (right) after LPC incorporation into the liposome patch. (C) The activation of the MscL mutant channel in the presence of 5 μM LPC in the patch pipette before, during and after application of suction to the pipette. Note longer openings of multiple channels when compared to 3 μM LPC recordings. Pipette potential was +30 mV.