Development of systems for time-resolved intermolecular interaction detection during protein reactions and application to reaction mechanism

2016 Fiscal Year Final Research Report

• Project/Area Number: 25288005
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Physical chemistry
• Research Institution: Kyoto University
• Principal Investigator: Terazima Masahide 京都大学, 理学(系)研究科(研究院), 教授 (00188674)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: タンパク質 / 反応 / 時間分解 / 拡散係数

Outline of Final Research Achievements

In order to measure the translational diffusion coefficient change by the transient grating method combined with a stopped flow system, we constructed a new stopped flow system for minimizing turbulence after mixing two solutions. This system can suppress the turbulence within 100 ms after the mixing. Furthermore, we improved the system to decrease a volume of required sample solution, and succeeded in mixing only a few micro liter solution, which is a big advantage for application to protein samples. By using this system, we measured a diffusion coefficient change in time-domain after acid denaturation of a photosensor protein of phototropin sample. Moreover, association reaction of a protein of KaiC which has a function to control the circadian rhythm was detected in time-domain.

Free Research Field

生物物理化学