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. 2018 Oct 1;9:1389. doi: 10.3389/fphys.2018.01389

FIGURE 5.

FIGURE 5

Gating alteration in the triple mutant L109S/N117C/N167C MscS. (A) Growth curves of MJF465 cells expressing WT, N117C/N167C, L109S, and L109S/N117C/N167C MscS induced with IPTG (circles) or without induction (triangles). Arrows indicate induction by 1 mM IPTG. (B) Representative single-channel recordings of MscS expressed in the E. coli MJF465 strain, from the same patch before and after treatment with copper phenanthroline [Cu(II)]. Lower traces: applied negative pressure. (C) Analysis of maximum current amplitude before and after treatment with Cu(II) (mean ± SEM, n = 6). (D) Representative single-channel recordings of L109S/N117C/N167C, from the same patch before and after treatment with βME. Lower traces: applied negative pressure. (E) Single-channel openings of L109S and L109S/N117C/N167C MscS (left; dashed horizontal lines indicate the closed state) and count vs. amplitude histograms (right). (F) Conductance of L109S and L109S/N117C/N167C MscS determined at 20 mV membrane potential (mean ± SEM, n = 6).