Disulfide mapping the voltage-sensing mechanism of a voltage-dependent potassium channel

Tomohiro Nozaki, Shin Ichiro Ozawa, Hitomi Harada, Tomomi Kimura, Masanori Osawa, Ichio Shimada

Research output: Contribution to journalArticle

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Abstract

Voltage-dependent potassium (Kv) channels allow for the selective permeability of potassium ions in a membrane potential dependent manner, playing crucial roles in neurotransmission and muscle contraction. Kv channel is a tetramer, in which each subunit possesses a voltage-sensing domain (VSD) and a pore domain (PD). Although several lines of evidence indicated that membrane depolarization is sensed as the movement of helix S4 of the VSD, the detailed voltage-sensing mechanism remained elusive, due to the difficulty of structural analyses at resting potential. In this study, we conducted a comprehensive disulfide locking analysis of the VSD using 36 double Cys mutants, in order to identify the proximal residue pairs of the VSD in the presence or absence of a membrane potential. An intramolecular SS-bond was formed between 6 Cys pairs under both polarized and depolarized environment, and one pair only under depolarized environment. The multiple conformations captured by the SS-bond can be divided by two states, up and down, where S4 lies on the extracellular and intracellular sides of the membrane, respectively, with axial rotation of 180°. The transition between these two states is caused by the S4 translocation of 12 Å, enabling allosteric regulation of the gating at the PD.

Original languageEnglish
Article number37303
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 2016 Nov 17

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Potassium Channels
Disulfides
Membrane Potentials
Allosteric Regulation
Voltage-Gated Potassium Channels
Intracellular Membranes
Muscle Contraction
Synaptic Transmission
Permeability
Potassium
Ions
Membranes

ASJC Scopus subject areas

  • General

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Disulfide mapping the voltage-sensing mechanism of a voltage-dependent potassium channel. / Nozaki, Tomohiro; Ozawa, Shin Ichiro; Harada, Hitomi; Kimura, Tomomi; Osawa, Masanori; Shimada, Ichio.

In: Scientific Reports, Vol. 6, 37303, 17.11.2016.

Research output: Contribution to journalArticle

Nozaki, Tomohiro ; Ozawa, Shin Ichiro ; Harada, Hitomi ; Kimura, Tomomi ; Osawa, Masanori ; Shimada, Ichio. / Disulfide mapping the voltage-sensing mechanism of a voltage-dependent potassium channel. In: Scientific Reports. 2016 ; Vol. 6.
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