The hydrophobic nature of a novel membrane interface regulates the enzyme activity of a voltage-sensing phosphatase

Abstract
Data availability
Article and author information
Metrics

Abstract

Voltage-sensing phosphatases (VSP) contain a voltage sensor domain (VSD) similar to that of voltage-gated ion channels but lack a pore-gate domain. A VSD in a VSP regulates the cytoplasmic catalytic region (CCR). However, the mechanisms by which the VSD couples to the CCR remain elusive. Here we report a membrane interface (named 'the hydrophobic spine'), which is essential for the coupling of the VSD and CCR. Our molecular dynamics simulations suggest that the hydrophobic spine of Ciona intestinalis VSP (Ci-VSP) provides a hinge-like motion for the CCR through the loose membrane association of the phosphatase domain. Electrophysiological experiments indicate that the voltage-dependent phosphatase activity of Ci-VSP depends on the hydrophobicity and presence of an aromatic ring in the hydrophobic spine. Analysis of conformational changes in the VSD and CCR suggests that the VSP has two states with distinct enzyme activities and that the second transition depends on the hydrophobic spine.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 4,5,6,7,8,9 and 10.

Article and author information

Author details

Akira Kawanabe

Department of Physiology, Graduate School of Medicine, Osaka University, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Masaki Hashimoto

Graduate School of Frontier Biosciences, Osaka University, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Manami Nishizawa

School of Medical Technology, Teikyo University, Tokyo, Japan

Competing interests
The authors declare that no competing interests exist.
Kazuhisa Nishizawa

School of Medical Technology, Teikyo University, Tokyo, Japan

Competing interests
The authors declare that no competing interests exist.
Hirotaka Narita

Institute for Protein Research, Osaka University, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Tomoko Yonezawa

Department of Physiology, Graduate School of Medicine, Osaka University, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Yuka Jinno

Department of Physiology, Graduate School of Medicine, Osaka University, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Souhei Sakata

Department of Physiology, Division of Life Sciences, Faculty of Medicine, Osaka Medical College, Takatsuki, Japan

Competing interests
The authors declare that no competing interests exist.
Atsushi Nakagawa

Institute for Protein Research, Osaka University, Suita, Japan

Competing interests
The authors declare that no competing interests exist.
Yasushi Okamura

Department of Physiology, Graduate School of Medicine, Osaka University, Suita, Japan

For correspondence
yokamura@phys2.med.osaka-u.ac.jp

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-5386-7968

Funding

Core Research for Evolutional Science and Technology (JPMJCR14M3)

Atsushi Nakagawa
Yasushi Okamura

Ministry of Education, Culture, Sports, Science, and Technology (25253016)

Yasushi Okamura

Japan Society for the Promotion of Science (15H05901)

Yasushi Okamura

Japan Society for the Promotion of Science (JP15K18516)

Akira Kawanabe

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All experiments were carried out following the guidelines of the Animal Research Committees of the Graduate School of Medicine of Osaka University.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.