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Engineering voltage sensing phosphatase (VSP).

Hidekazu Tsutsui1, Natsuki Mizutani2, Yasushi Okamura2

  • 1School of Materials Science, Japan Advanced Institute of Science and Technology (JAIST), Nomi, Ishikawa, Japan.

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Voltage sensing phosphatase (VSP) links voltage sensing to enzyme activity. Its voltage sensor domain is a versatile tool for creating voltage indicators and manipulating cellular phosphoinositide levels.

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Gene-encoded voltage indicatorIon channelMembrane potentialPIP2Phosphoinositide

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Cell Biology

Background:

  • Voltage sensing phosphatase (VSP) integrates a voltage sensor domain (VSD) with a phosphoinositide (PIP) phosphatase region.
  • The VSD shares structural similarities with voltage-gated ion channels, while the phosphatase region resembles the tumor suppressor enzyme PTEN.
  • VSP's activity is activated by membrane depolarization through a coupled mechanism between the VSD and the enzyme domain.

Purpose of the Study:

  • To explore the unique properties and applications of Voltage sensing phosphatase (VSP).
  • To highlight the VSD's potential as a transferable module for voltage sensitivity.
  • To discuss VSP's utility in altering cellular PIP levels and its role in developing novel molecular tools.

Main Methods:

  • Investigating the structure-function relationship of VSP.
  • Utilizing the VSD of VSP to engineer gene-encoded voltage indicators (GEVIs).
  • Employing VSP for acute and transient reduction of cellular PIP levels, specifically PI(4,5)P2, via voltage protocols.

Main Results:

  • Demonstrated that the VSD of VSP is a self-contained module capable of conferring voltage sensitivity to other proteins.
  • Successfully developed various GEVIs by combining the VSP VSD with fluorescent proteins.
  • Showcased VSP's ability to acutely reduce cellular PIP levels upon application of a voltage protocol.

Conclusions:

  • VSP is a powerful tool for both voltage sensing and modulating intracellular phosphoinositide dynamics.
  • The modular nature of the VSP VSD enables versatile applications in biosensing and cell biology.
  • Advances in understanding VSP's molecular mechanisms facilitate its optimization as a research tool.