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The cytoplasmic coiled-coil mediates cooperative gating temperature sensitivity in the voltage-gated H+ channel Hv1

Yuichiro Fujiwara (), Tatsuki Kurokawa, Kohei Takeshita, Megumi Kobayashi, Yoshifumi Okochi, Atsushi Nakagawa and Yasushi Okamura ()
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Yuichiro Fujiwara: Laboratory of Integrative Physiology, Graduate School and Faculty of Medicine, Osaka University, Yamadaoka 2-2, Suita
Tatsuki Kurokawa: Laboratory of Integrative Physiology, Graduate School and Faculty of Medicine, Osaka University, Yamadaoka 2-2, Suita
Kohei Takeshita: Laboratory of Supramolecular Crystallography, Research Center for State-of-the-Art Functional Protein Analysis, Institute for Protein Research, Osaka University, Yamadaoka 3-2, Suita, Osaka 565-0871, Japan.
Megumi Kobayashi: Laboratory of Integrative Physiology, Graduate School and Faculty of Medicine, Osaka University, Yamadaoka 2-2, Suita
Yoshifumi Okochi: Laboratory of Integrative Physiology, Graduate School and Faculty of Medicine, Osaka University, Yamadaoka 2-2, Suita
Atsushi Nakagawa: Laboratory of Supramolecular Crystallography, Research Center for State-of-the-Art Functional Protein Analysis, Institute for Protein Research, Osaka University, Yamadaoka 3-2, Suita, Osaka 565-0871, Japan.
Yasushi Okamura: Laboratory of Integrative Physiology, Graduate School and Faculty of Medicine, Osaka University, Yamadaoka 2-2, Suita

Nature Communications, 2012, vol. 3, issue 1, 1-11

Abstract: Abstract Hv1/VSOP is a dimeric voltage-gated H+ channel in which the gating of one subunit is reportedly coupled to that of the other subunit within the dimer. The molecular basis for dimer formation and intersubunit coupling, however, remains unknown. Here we show that the carboxy terminus ends downstream of the S4 voltage-sensor helix twist in a dimer coiled-coil architecture, which mediates cooperative gating. We also show that the temperature-dependent activation of H+ current through Hv1/VSOP is regulated by thermostability of the coiled-coil domain, and that this regulation is altered by mutation of the linker between S4 and the coiled-coil. Cooperative gating within the dimer is also dependent on the linker structure, which circular dichroism spectrum analysis suggests is α-helical. Our results indicate that the cytoplasmic coiled-coil strands form continuous α-helices with S4 and mediate cooperative gating to adjust the range of temperatures over which Hv1/VSOP operates.

Date: 2012
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DOI: 10.1038/ncomms1823

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