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. 2020 Jul 23;11:3690. doi: 10.1038/s41467-020-17538-1

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© The Author(s) 2020

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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Fig. 3 — a, b Orthogonal views of TM5. Glycine and alanine residues are indicated as red spheres. A320 and its contacts in a neighboring helix (A322 and L329) are highlighted in green. V319 and F323 constitute the hydrophobic gate. c Representative traces for single AtMSL1 and A320V channel activities in response to a brief 1 s pressure ramp followed by 90 s without pressure. Closed state (baseline) current is indicated by C while open states are indicated by O. The maximum negative pressure used is indicated. Membrane potential was clamped at −70 mV. Multiple subconducting states were observed with the A320V mutant, while the peak unitary current of A320V was indistinguishable from that of the wild type (Table 3). d Percentages of AtMSL1 (black) and AtMSL1 A320V (gray) gating events with open dwell times of 0–19.99 s, 20–39.99 s, 40–59.99 s, 60–79.99 s, or 80+ s. Data represent traces obtained from 10 patches per variant channel with only the first gating event analyzed per trace (103 traces for AtMSL1, 34 for AtMSL1 A320V). Source data are provided as a Source Data file.