| Co-Investigator(Kenkyū-buntansha) |
中野 辰彦 日本バイオラッドラボラトリーズ(株), 分析機器, 課長
MOGI Tatsushi Tokyo Univ., Science, Instructor, 大学院・理学研究科, 助手 (90219965)
TASUHIKO Nakano Bio-Rad, Analystical Lab, Manager
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| Research Abstract |
This research project aims at constructing a new Fourier-transform infrared (FTIR) spectrometer, which can probe protein structural changes for light-induced irreversible chemical reactions at room temperature.
Time-resolved step-scan FTIR spectroscopy is a powerful tool in investigation of light-induced chemical reactions in biological molecules, and extensive studies have been indeed conducted for bacteriorhodopsin, a light-driven proton pump. However, the system cannot be applied to visual rhodopsin, because more than 1000 laser illuminations are necessary to obtain time-resolved infrared spectra and visual rhodopsin easily bleaches upon light illumination. Thus, only cyclic reactions are applicable for current time-resolved step-scan FTIR spectrometer, and no information can be obtained for irreversible reactions.
In this project, we combined time-resolved step-scan FTIR spectrometer and IR microscope that contains movable xy stage. In this system, sample on the xy stage is synchronously moved to laser illumination, so that new sample is supplied for every illumination. During the project period, several problems have been settled to optimize the new system, and it will provide new information on biophysical aspects in functional processes of proteins. The system can be also applied to oxidases, because light-induced reduction of this enzyme initiates reaction.
While we constructed the new FTIR spectrometer, we obtained various results on structure-function relationship in rhodopsins and oxidases, as shown in the publication list. Such new information will lead to better understanding of molecular mechanism in bioenergetics.
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