Analysis of rotational motion of voltage sensor domain based on phosphorescence polarization

2017 Fiscal Year Final Research Report

• Project/Area Number: 26670103
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General physiology
• Research Institution: Japan Advanced Institute of Science and Technology
• Principal Investigator: TSUTSUI Hidekazu 北陸先端科学技術大学院大学, 先端科学技術研究科, 准教授 (30392038)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Keywords: 電位センサードメイン / 燐光偏光 / 膜電位固定

Outline of Final Research Achievements

To investigate the transition mechanism of the voltage-sensor domain, an experimental setup for detecting segment rotation using phosphorescence polarization was built. The reliable quantitative measurements were not reached due to the two major problems that the membrane becomes remarkably fragile under the low oxygen condition required for the phosphorescence measurement and that it is difficult to correctly estimate the contribution of non-specific binding of phosphorescence dye to membrane. In the future, there is a possibility that it can be dealt with by using a brighter phosphorescent dye or a highly specific labeling technique that reduces the background light by using a total reflection illumination system. In the final year of the research period, we showed that the potential sensor performs global structural change by using environmentally responsive unnatural amino acid. We also clarified some unique properties of divalent cation permeability of CatSper voltage-sensor.

Free Research Field

生理学