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. 2019 May 24;12:133. doi: 10.3389/fnmol.2019.00133

Figure 7.

Figure 7

Conserved effects of mutations in the palm. All experiments of this figure are from hASIC1a expressed in Xenopus oocytes, voltage-clamped to −60 mV. (A,B) Overview and detailed image of a structural model of hASIC1a, based on the crystal structure of cASIC1a (Baconguis et al., 2014). The domains of one subunit are indicated by different colors, as in Figure 1A. (C) Left, representative current traces of the mutant Q276C in the background of hASIC1a-D212 (top) or -G212 (bottom), as indicated, before and after MTSET exposure (1 mM, 3 min). ASIC currents were induced by changing from the conditioning pH7.4 to the stimulation pH5.0. Right, the Isust/Ipeak ratio at pH5.0 (n = 8–10) is shown. For Q276C D212C, the Isust/Ipeak ratio was measured during the last 2 s of the acid perfusion; the duration of the pH5.0 perfusion was the same in the control and MTSET condition. For Q276C G212, this ratio was measured at 16–18 s after the start of the pH5.0 perfusion. (D) SSD pH dependence of the mutant E418C before (open symbols) or after MTSET exposure (filled symbols, 1 mM 3 min, n = 4–7). Normalized current amplitudes are plotted as function of the conditioning pH. Two-way ANOVA showed that MTSET induced an acidic shift of the pH dependence in both backgrounds (p < 0.0001). (E) Left, representative current traces of the mutant N416C in the background of hASIC1a-D212 (top) or -G212 (bottom), before and after MTSET exposure (1 mM, 3 min). ASIC currents were induced by changing from the conditioning pH7.4 to the stimulation pH5.0. Right, the time constant of the current decay measured at pH 5.0, is shown before and after exposure to MTSET (n = 5–6). For (C,E), ***p < 0.001; ****p < 0.0001; paired t-test.