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. 2018 May 7;150(5):731–750. doi: 10.1085/jgp.201711965

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© 2018 Silverå Ejneby et al.

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Figure 9. — The role of S4 charges for the effect of Wu161 and other DHAA derivatives. (A) Schematic picture of mutated S4 charges on the Shaker K_V channel. The top gating charge (an arginine, R362 in WT) was moved step-by-step further out on S4 (left), or removed (R362Q; right). (B) Wu161-induced G(V) shifts on the Shaker K_V channel S4 arginine mutants. 100 µM, pH = 7.4. Mean ± SEM (n = 3–5). Shifts are compared with R362Q (dashed line), one-way ANOVA with Dunnett’s multiple comparison test. Blue: nonsignificant, P > 0.05. Green: larger effect than for R362Q, P < 0.001. Orange: smaller effect than for R362Q, P < 0.001. Data for the mutations in control solution at pH 7.4 according to Eq. 2: R362Q (V_½ = 3.4 ± 2.9 mV, s = 12.0 ± 0.8 mV, n = 4), R362 (WT) (V_½ = −21.4 ± 1.8 mV, s = 10.8 ± 1.1 mV, n = 3), L361R/R362Q (V_½ = −50.5 ± 1.4 mV, s = 17.9 ± 0.3 mV, n = 4), I360R/R362Q (V_½ = 24.6 ± 2.3 mV, s = 12.2 ± 1.0 mV, n = 5), A359R/R362Q (V_½ = 32.3 ± 3.8 mV, s = 13.0 ± 1.8 mV, n = 4), L358R/R362Q (V_½ = −12.1 ± 0.9 mV, s = 11.6 ± 0.3 mV, n = 4), S357R/R362Q (V_½ = −34.7 ± 2.1 mV, s = 8.1 ± 0.2 mV, n = 4), M356R/R362Q (V_½ = 19.3 ± 2.5 mV, s = 9.0 ± 0.1 mV, n = 4), and A359R/M356R (3R; V_½ = 25.4 ± 2.0 mV, s = 12.3 ± 0.9 mV, n = 6). (C) Correlation between G(V) shifts for the PUFA DHA (Ottosson et al., 2014) and for Wu161 on the Shaker K_V channel S4 arginine mutants (100 µM, pH = 7.4). Error bars represent mean ± SEM (n = 4–14). Slope (2.1 ± 0.2) is significantly different from 0 (Pearson correlation test and linear regression are both significant). (D) Effects of DHAA(p1), Wu32(p1) (Ottosson et al., 2017), Wu161(p3), and PUFA DHA (Ottosson et al., 2014) on the R362Q, WT, and 3R Shaker K_V channels (100 µM, pH = 7.4). Error bars represent mean ± SEM (n = 4–15).