Kinetic analyses of the local viscosity of the hydration layer of actomyosin at work studied by frequency-domain fluorescence polarization
Project/Area Number |
24570178
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Biophysics
|
Research Institution | Osaka University (2014-2015) Tohoku University (2012-2013) |
Principal Investigator |
|
Project Period (FY) |
2012-04-01 – 2016-03-31
|
Project Status |
Completed (Fiscal Year 2015)
|
Budget Amount *help |
¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2012: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
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Keywords | アクチン / 周波数蛍光法 / 回転ブラウン運動 / 水和 / wobbling-in-coneモデル / 蛍光異方性 / 時間分解蛍光法 / rhodamine 6G / 蛍光偏光 / 回転拡散 / 周波数領域蛍光 / オリゴエチレングリコール / 局所粘度 / wobbling-in-cone model / 周波数領域蛍光法 / 時間分解蛍光偏光 / フィラメント化 / 局所粘性 / 回転相関時間 / 周波数領域蛍光測定 / 蛍光寿命 / LN光変調器 / 蛍光偏光解消法 / アクトミオシン / キネティクス |
Outline of Final Research Achievements |
Hydration water is closely involved in the stability and dynamics of protein and plays a vital role in the mechanism of protein function. Here we develop a technique to analyse physical properties of the hydration water around protein and apply this technique to actin, a muscle contractile protein. A fluorescent dye is conjugated to actin through a flexible linker so that the dye is tethered in the hydration layer, and its time-resolved fluorescence polarisation is measured by a newly in-house built high-precision frequency-domain fluorometer that uses an LN light modulator to generate modulated excitation light. The dye undergoes rotational Brownian motion in the hydration layer of actin that can be analysed by the measurement, and the analysis reveals a hydrodynamic property of the hydration layer around actin. The technique developed in this study is useful to move a step closer to understanding the role of hydration water around protein.
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Report
(5 results)
Research Products
(18 results)