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. 2005 Dec;126(6):605–617. doi: 10.1085/jgp.200509413

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Copyright © 2005, The Rockefeller University Press

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Figure 9. — The two different gating modes of the AKT2 wild type and the mutant AKT2-K197S. (A–F) Representative K⁺ currents obtained from COS cells expressing AKT2, AKT2-S210A-S329A, AKT2-S210N-S329N, AKT2-K197S, AKT2-K197S-S210A-S329A, and AKT2-K197S-S210N-S329N. Pulse protocols: currents elicited by voltage steps (duration T) from a holding potential of V_H to voltages from V_min to V_max (20-mV increments) followed by a further voltage step to V_T (pulse protocol indicated as [V_H; V_min … V_max, T; V_T] in the following). The dashed lines indicate the zero current level. (A) AKT2; (+40 mV; −140 mV … +140 mV, 3 s; +40 mV). (B) The mutant S210A-S329A mimics AKT2-mode#1 gating (+40 mV; −140 mV … +40 mV, 3 s; +40 mV). (C) The mutant S210N-S329N mimics AKT2-mode#2 gating (+40 mV; −140 mV … +140 mV, 2 s; +40 mV). (D) AKT2-K197S; (0 mV; −160 mV … +40 mV, 1.5 s; −80 mV). (E) The mutant K197S-S210A-S329A mimics AKT2-K197S-mode#1 gating (0 mV; −200 mV … +40 mV, 1.5 s; 0 mV). (F) The mutant K197S-S210N-S329N mimics AKT2-K197S-mode#2 gating (0 mV; −180 mV … +40 mV, 1.5 s; −80 mV). (G) Relative conductance of AKT2 (black diamonds), AKT2-S210A-S329A (black circles), and AKT2-S210N-S329N (black triangles). Data are displayed as mean ± SD (n = 3–6). Gray lines represent nonlinear least-squares fittings of Eq. 4 to the data for S210A-S329A and S210N-S329N (R² > 0.95). Fits were constrained by the adjustment of a common value δ for both datasets, yielding δ = 0.78, V_1/2 ^S210A-S329A = −151.9 mV, V_1/2 ^S210N-S329N = +136.0 mV. The solid line represents a weighted sum of the two gray lines: fit_rel.G_AKT2 = 0.56 × fit_rel.G_S210A-S329A + 0.44 × fit_rel.G_S210N-S329N. (H) Relative conductance of AKT2-K197S (gray diamonds), AKT2-K197S-S210A-S329A (gray circles), and AKT2-K197S-S210N-S329N (gray triangles). Data are displayed as mean ± SD (n = 3–6). Gray lines represent nonlinear least-squares fittings of Eq. 4 to the data for K197S-S210A-S329A and K197S-S210N-S329N (R² > 0.95). Fits were constrained by the adjustment of a common value δ for both datasets, yielding δ = 1.22, V_1/2 ^{K197S-S210A-S329A} = −183.1 mV, V_1/2 ^{K197S-S210N-S329N} = −135.5 mV. The solid line represents a weighted sum of the two gray lines: fit_rel.G_K197S = 0.46 × fit_rel.G_{K197S-S210A-S329A} + 0.54 × fit_rel.G_{K197S-S210N-S329N}.