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Domain-specific charge neutralizations. (A) Sequence alignments of the S4 voltage-sensing segments from each domain in Nav1.4. Mutations are marked in bold. (B) A representative family of ionic current traces for wild-type and mutant channels in response to 30-ms depolarizing pulses from −110 to 65 mV (10-mV steps), preceded by a 20-ms prepulse to −120 mV from a holding potential of −80 mV. (C) Normalized peak G-V relationship for wild-type and mutant channels. Solid lines are single <t>Boltzmann</t> fits to the mean for each construct (wild type: V 1/2 = −23.8 mV and z = 5.8 e − ; DI-CN: V 1/2 = −36.5 mV and z = 3.8 e − ; DII-CN: V 1/2 = −25.1 mV and z = 3.3 e − ; DIII-CN: V 1/2 = −26.4 mV and z = 3.9 e − ; DIV-CN: V 1/2 = −23.1 mV and z = 3.8 e − ).
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Domain-specific charge neutralizations. (A) Sequence alignments of the S4 voltage-sensing segments from each domain in Nav1.4. Mutations are marked in bold. (B) A representative family of ionic current traces for wild-type and mutant channels in response to 30-ms depolarizing pulses from −110 to 65 mV (10-mV steps), preceded by a 20-ms prepulse to −120 mV from a holding potential of −80 mV. (C) Normalized peak G-V relationship for wild-type and mutant channels. Solid lines are single <t>Boltzmann</t> fits to the mean for each construct (wild type: V 1/2 = −23.8 mV and z = 5.8 e − ; DI-CN: V 1/2 = −36.5 mV and z = 3.8 e − ; DII-CN: V 1/2 = −25.1 mV and z = 3.3 e − ; DIII-CN: V 1/2 = −26.4 mV and z = 3.9 e − ; DIV-CN: V 1/2 = −23.1 mV and z = 3.8 e − ).
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lattice boltzmann method  (Lallemand inc)
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Domain-specific charge neutralizations. (A) Sequence alignments of the S4 voltage-sensing segments from each domain in Nav1.4. Mutations are marked in bold. (B) A representative family of ionic current traces for wild-type and mutant channels in response to 30-ms depolarizing pulses from −110 to 65 mV (10-mV steps), preceded by a 20-ms prepulse to −120 mV from a holding potential of −80 mV. (C) Normalized peak G-V relationship for wild-type and mutant channels. Solid lines are single <t>Boltzmann</t> fits to the mean for each construct (wild type: V 1/2 = −23.8 mV and z = 5.8 e − ; DI-CN: V 1/2 = −36.5 mV and z = 3.8 e − ; DII-CN: V 1/2 = −25.1 mV and z = 3.3 e − ; DIII-CN: V 1/2 = −26.4 mV and z = 3.9 e − ; DIV-CN: V 1/2 = −23.1 mV and z = 3.8 e − ).
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Domain-specific charge neutralizations. (A) Sequence alignments of the S4 voltage-sensing segments from each domain in Nav1.4. Mutations are marked in bold. (B) A representative family of ionic current traces for wild-type and mutant channels in response to 30-ms depolarizing pulses from −110 to 65 mV (10-mV steps), preceded by a 20-ms prepulse to −120 mV from a holding potential of −80 mV. (C) Normalized peak G-V relationship for wild-type and mutant channels. Solid lines are single Boltzmann fits to the mean for each construct (wild type: V 1/2 = −23.8 mV and z = 5.8 e − ; DI-CN: V 1/2 = −36.5 mV and z = 3.8 e − ; DII-CN: V 1/2 = −25.1 mV and z = 3.3 e − ; DIII-CN: V 1/2 = −26.4 mV and z = 3.9 e − ; DIV-CN: V 1/2 = −23.1 mV and z = 3.8 e − ).

Journal: The Journal of General Physiology

Article Title: Domain IV voltage-sensor movement is both sufficient and rate limiting for fast inactivation in sodium channels

doi: 10.1085/jgp.201310998

Figure Lengend Snippet: Domain-specific charge neutralizations. (A) Sequence alignments of the S4 voltage-sensing segments from each domain in Nav1.4. Mutations are marked in bold. (B) A representative family of ionic current traces for wild-type and mutant channels in response to 30-ms depolarizing pulses from −110 to 65 mV (10-mV steps), preceded by a 20-ms prepulse to −120 mV from a holding potential of −80 mV. (C) Normalized peak G-V relationship for wild-type and mutant channels. Solid lines are single Boltzmann fits to the mean for each construct (wild type: V 1/2 = −23.8 mV and z = 5.8 e − ; DI-CN: V 1/2 = −36.5 mV and z = 3.8 e − ; DII-CN: V 1/2 = −25.1 mV and z = 3.3 e − ; DIII-CN: V 1/2 = −26.4 mV and z = 3.9 e − ; DIV-CN: V 1/2 = −23.1 mV and z = 3.8 e − ).

Article Snippet: Boltzmann fits to conductance–voltage relations were performed in Origin (OriginLab).

Techniques: Sequencing, Mutagenesis, Construct

DIV voltage-sensor movement is sufficient for fast inactivation. (A) A representative family of current traces in response to a 30-ms test pulse to −30 mV to assay the fraction of non–steady-state inactivated channels after a 100-ms conditioning prepulse between −170 and 50 mV. See Results for a detailed description of the voltage-pulse protocol. (B) Normalized steady-state inactivation versus conditioning prepulse voltage for wild-type and mutant channels. Solid lines are single Boltzmann fits to the mean for each construct (wild type: V 1/2 = −62.3 mV and z = 4.7 e − ; DI-CN: V 1/2 = −69.2 mV and z = 4.2 e − ; DII-CN: V 1/2 = −65.8 mV and z = 3.7 e − ; DIII-CN: V 1/2 = −68.7 mV and z = 2.7 e − ; DIV-CN: V 1/2 = −137.3 mV and z = 1.1 e − ). Note that the relation does not saturate above −200 mV for DIV-CN, so the observed left-shift in the steady-state inactivation versus voltage relation reflects the minimum shift conferred by this mutant.

Journal: The Journal of General Physiology

Article Title: Domain IV voltage-sensor movement is both sufficient and rate limiting for fast inactivation in sodium channels

doi: 10.1085/jgp.201310998

Figure Lengend Snippet: DIV voltage-sensor movement is sufficient for fast inactivation. (A) A representative family of current traces in response to a 30-ms test pulse to −30 mV to assay the fraction of non–steady-state inactivated channels after a 100-ms conditioning prepulse between −170 and 50 mV. See Results for a detailed description of the voltage-pulse protocol. (B) Normalized steady-state inactivation versus conditioning prepulse voltage for wild-type and mutant channels. Solid lines are single Boltzmann fits to the mean for each construct (wild type: V 1/2 = −62.3 mV and z = 4.7 e − ; DI-CN: V 1/2 = −69.2 mV and z = 4.2 e − ; DII-CN: V 1/2 = −65.8 mV and z = 3.7 e − ; DIII-CN: V 1/2 = −68.7 mV and z = 2.7 e − ; DIV-CN: V 1/2 = −137.3 mV and z = 1.1 e − ). Note that the relation does not saturate above −200 mV for DIV-CN, so the observed left-shift in the steady-state inactivation versus voltage relation reflects the minimum shift conferred by this mutant.

Article Snippet: Boltzmann fits to conductance–voltage relations were performed in Origin (OriginLab).

Techniques: Mutagenesis, Construct