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Allosteric voltage gating of potassium channels I. Mslo ionic currents in the absence of Ca(2+).

Horrigan FT, Cui J, Aldrich RW - J. Gen. Physiol. (1999)

Bottom Line: However, the time constant of I(K) relaxation [tau(I(K))] exhibits a complex voltage dependence that is inconsistent with models that contain a single rate limiting step. tau(I(K)) increases weakly with voltage from -500 to -20 mV, with an equivalent charge (z) of only 0.14 e, and displays a stronger voltage dependence from +30 to +140 mV (z = 0.49 e), which then decreases from +180 to +240 mV (z = -0.29 e).These results can be understood in terms of a gating scheme where a central transition between a closed and an open conformation is allosterically regulated by the state of four independent and identical voltage sensors.These conclusions not only provide a framework for interpreting studies of large conductance Ca(2+)-activated K(+) channel voltage gating, but also have important implications for understanding the mechanism of Ca(2+) sensitivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA.

ABSTRACT
Activation of large conductance Ca(2+)-activated K(+) channels is controlled by both cytoplasmic Ca(2+) and membrane potential. To study the mechanism of voltage-dependent gating, we examined mSlo Ca(2+)-activated K(+) currents in excised macropatches from Xenopus oocytes in the virtual absence of Ca(2+) (<1 nM). In response to a voltage step, I(K) activates with an exponential time course, following a brief delay. The delay suggests that rapid transitions precede channel opening. The later exponential time course suggests that activation also involves a slower rate-limiting step. However, the time constant of I(K) relaxation [tau(I(K))] exhibits a complex voltage dependence that is inconsistent with models that contain a single rate limiting step. tau(I(K)) increases weakly with voltage from -500 to -20 mV, with an equivalent charge (z) of only 0.14 e, and displays a stronger voltage dependence from +30 to +140 mV (z = 0.49 e), which then decreases from +180 to +240 mV (z = -0.29 e). Similarly, the steady state G(K)-V relationship exhibits a maximum voltage dependence (z = 2 e) from 0 to +100 mV, and is weakly voltage dependent (z congruent with 0.4 e) at more negative voltages, where P(o) = 10(-5)-10(-6). These results can be understood in terms of a gating scheme where a central transition between a closed and an open conformation is allosterically regulated by the state of four independent and identical voltage sensors. In the absence of Ca(2+), this allosteric mechanism results in a gating scheme with five closed (C) and five open (O) states, where the majority of the channel's voltage dependence results from rapid C-C and O-O transitions, whereas the C-O transitions are rate limiting and weakly voltage dependent. These conclusions not only provide a framework for interpreting studies of large conductance Ca(2+)-activated K(+) channel voltage gating, but also have important implications for understanding the mechanism of Ca(2+) sensitivity.

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Properties of the allosteric voltage-gating scheme. (A) The τ(IK)–V relationship determined by simulating Fig. 9 (•; Table : average 5°C) can be reproduced by an analytical approximation (solid line) that assumes horizontal transitions are equilibrated. The voltage dependence of time constants for individual C–O transitions are also plotted (τi = [δi + γi)−1]. (B) Po–V relationships predicted by Fig. 9 (solid lines) are plotted on a semi-log scale as the allosteric factor D is adjusted [with zL = 0.4 e, zJ = 0.55 e, Vh(J) = 145]. The equilibrium constant L was adjusted together with D such that the half-activation voltage remained constant (for D = 5–160: L = 2.18 e−4, 1.57 e−5, 1.05 e−6, 6.80 e−8, 4.33 e−9, and 2.72 e−10). A dashed line indicates the prediction of sequential Fig. 5 (from Fig. 6).
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Figure 10: Properties of the allosteric voltage-gating scheme. (A) The τ(IK)–V relationship determined by simulating Fig. 9 (•; Table : average 5°C) can be reproduced by an analytical approximation (solid line) that assumes horizontal transitions are equilibrated. The voltage dependence of time constants for individual C–O transitions are also plotted (τi = [δi + γi)−1]. (B) Po–V relationships predicted by Fig. 9 (solid lines) are plotted on a semi-log scale as the allosteric factor D is adjusted [with zL = 0.4 e, zJ = 0.55 e, Vh(J) = 145]. The equilibrium constant L was adjusted together with D such that the half-activation voltage remained constant (for D = 5–160: L = 2.18 e−4, 1.57 e−5, 1.05 e−6, 6.80 e−8, 4.33 e−9, and 2.72 e−10). A dashed line indicates the prediction of sequential Fig. 5 (from Fig. 6).

Mentions: According to Fig. 9, the horizontal C–C and O–O transitions will be so fast that they do not affect τ(IK). This point was confirmed in Fig. 10 A by showing that the values of τ(IK) measured from IK simulations (symbols) can be reproduced by an analytical approximation of the τ(IK)–V relationship (solid line) that assumes horizontal transitions are equilibrated: 18\documentclass[10pt]{article}\usepackage{amsmath}\usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy}\usepackage{mathrsfs}\usepackage{pmc}\usepackage[Euler]{upgreek}\pagestyle{empty}\oddsidemargin -1.0in\begin{document}\begin{equation*}{\mathrm{{\tau}}} \left \left({\mathrm{I}}_{{\mathrm{K}}}\right) \right = \left \left[{{\sum_{{\mathrm{i}}}}} \left \left({\mathrm{{\delta}}}_{{\mathrm{i}}}p{\mathrm{C}}_{{\mathrm{i}}}+{\mathrm{{\gamma}}}_{{\mathrm{i}}}p{\mathrm{O}}_{{\mathrm{i}}}\right) \right \right] \right ^{-1}{\mathrm{,}}\end{equation*}\end{document} where δi and γi are rate constants for the Ci–Oi transitions and pCi and pOi are conditional occupancies of the open and closed states [pCi = p(Ci/C) and pOi = p(Oi/O)].


Allosteric voltage gating of potassium channels I. Mslo ionic currents in the absence of Ca(2+).

Horrigan FT, Cui J, Aldrich RW - J. Gen. Physiol. (1999)

Properties of the allosteric voltage-gating scheme. (A) The τ(IK)–V relationship determined by simulating Fig. 9 (•; Table : average 5°C) can be reproduced by an analytical approximation (solid line) that assumes horizontal transitions are equilibrated. The voltage dependence of time constants for individual C–O transitions are also plotted (τi = [δi + γi)−1]. (B) Po–V relationships predicted by Fig. 9 (solid lines) are plotted on a semi-log scale as the allosteric factor D is adjusted [with zL = 0.4 e, zJ = 0.55 e, Vh(J) = 145]. The equilibrium constant L was adjusted together with D such that the half-activation voltage remained constant (for D = 5–160: L = 2.18 e−4, 1.57 e−5, 1.05 e−6, 6.80 e−8, 4.33 e−9, and 2.72 e−10). A dashed line indicates the prediction of sequential Fig. 5 (from Fig. 6).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2230643&req=5

Figure 10: Properties of the allosteric voltage-gating scheme. (A) The τ(IK)–V relationship determined by simulating Fig. 9 (•; Table : average 5°C) can be reproduced by an analytical approximation (solid line) that assumes horizontal transitions are equilibrated. The voltage dependence of time constants for individual C–O transitions are also plotted (τi = [δi + γi)−1]. (B) Po–V relationships predicted by Fig. 9 (solid lines) are plotted on a semi-log scale as the allosteric factor D is adjusted [with zL = 0.4 e, zJ = 0.55 e, Vh(J) = 145]. The equilibrium constant L was adjusted together with D such that the half-activation voltage remained constant (for D = 5–160: L = 2.18 e−4, 1.57 e−5, 1.05 e−6, 6.80 e−8, 4.33 e−9, and 2.72 e−10). A dashed line indicates the prediction of sequential Fig. 5 (from Fig. 6).
Mentions: According to Fig. 9, the horizontal C–C and O–O transitions will be so fast that they do not affect τ(IK). This point was confirmed in Fig. 10 A by showing that the values of τ(IK) measured from IK simulations (symbols) can be reproduced by an analytical approximation of the τ(IK)–V relationship (solid line) that assumes horizontal transitions are equilibrated: 18\documentclass[10pt]{article}\usepackage{amsmath}\usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy}\usepackage{mathrsfs}\usepackage{pmc}\usepackage[Euler]{upgreek}\pagestyle{empty}\oddsidemargin -1.0in\begin{document}\begin{equation*}{\mathrm{{\tau}}} \left \left({\mathrm{I}}_{{\mathrm{K}}}\right) \right = \left \left[{{\sum_{{\mathrm{i}}}}} \left \left({\mathrm{{\delta}}}_{{\mathrm{i}}}p{\mathrm{C}}_{{\mathrm{i}}}+{\mathrm{{\gamma}}}_{{\mathrm{i}}}p{\mathrm{O}}_{{\mathrm{i}}}\right) \right \right] \right ^{-1}{\mathrm{,}}\end{equation*}\end{document} where δi and γi are rate constants for the Ci–Oi transitions and pCi and pOi are conditional occupancies of the open and closed states [pCi = p(Ci/C) and pOi = p(Oi/O)].

Bottom Line: However, the time constant of I(K) relaxation [tau(I(K))] exhibits a complex voltage dependence that is inconsistent with models that contain a single rate limiting step. tau(I(K)) increases weakly with voltage from -500 to -20 mV, with an equivalent charge (z) of only 0.14 e, and displays a stronger voltage dependence from +30 to +140 mV (z = 0.49 e), which then decreases from +180 to +240 mV (z = -0.29 e).These results can be understood in terms of a gating scheme where a central transition between a closed and an open conformation is allosterically regulated by the state of four independent and identical voltage sensors.These conclusions not only provide a framework for interpreting studies of large conductance Ca(2+)-activated K(+) channel voltage gating, but also have important implications for understanding the mechanism of Ca(2+) sensitivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California 94305, USA.

ABSTRACT
Activation of large conductance Ca(2+)-activated K(+) channels is controlled by both cytoplasmic Ca(2+) and membrane potential. To study the mechanism of voltage-dependent gating, we examined mSlo Ca(2+)-activated K(+) currents in excised macropatches from Xenopus oocytes in the virtual absence of Ca(2+) (<1 nM). In response to a voltage step, I(K) activates with an exponential time course, following a brief delay. The delay suggests that rapid transitions precede channel opening. The later exponential time course suggests that activation also involves a slower rate-limiting step. However, the time constant of I(K) relaxation [tau(I(K))] exhibits a complex voltage dependence that is inconsistent with models that contain a single rate limiting step. tau(I(K)) increases weakly with voltage from -500 to -20 mV, with an equivalent charge (z) of only 0.14 e, and displays a stronger voltage dependence from +30 to +140 mV (z = 0.49 e), which then decreases from +180 to +240 mV (z = -0.29 e). Similarly, the steady state G(K)-V relationship exhibits a maximum voltage dependence (z = 2 e) from 0 to +100 mV, and is weakly voltage dependent (z congruent with 0.4 e) at more negative voltages, where P(o) = 10(-5)-10(-6). These results can be understood in terms of a gating scheme where a central transition between a closed and an open conformation is allosterically regulated by the state of four independent and identical voltage sensors. In the absence of Ca(2+), this allosteric mechanism results in a gating scheme with five closed (C) and five open (O) states, where the majority of the channel's voltage dependence results from rapid C-C and O-O transitions, whereas the C-O transitions are rate limiting and weakly voltage dependent. These conclusions not only provide a framework for interpreting studies of large conductance Ca(2+)-activated K(+) channel voltage gating, but also have important implications for understanding the mechanism of Ca(2+) sensitivity.

Show MeSH
Related in: MedlinePlus