2021 Fiscal Year Research-status Report
Revealing oxidative stress in neurodegenerative brain by hydroxyl-radical-focused imaging and proteomics
| Project/Area Number |
21K14752
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| Research Institution | Kyoto University |
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Principal Investigator |
朱 浩 京都大学, 工学研究科, 助教 (90874545)
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| Project Period (FY) |
2021-04-01 – 2023-03-31
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| Keywords | oxidative stress / neurodegeneration / reactive oxygen species / chemical proteomics |
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| Outline of Annual Research Achievements |
The production of hydroxyl radical in our bodies is closely associated with the onset of many human diseases, due to its high reactivity that can cause severe damages on the functional biomolecules. This research aims at developing a new approach for the imaging- and proteomics-based analysis of hydroxyl radical in living systems. As planned, in the first stage, I focused on the development of chemical probes for hydroxyl radical-responsive protein labeling. A library having 10 members of phenol/aniline-derived probes have been synthesized, and their protein labeling behaviors in response to hydroxyl radical have been evaluated in test tubes. Certain substitutions on the phenyl ring could significantly improve the hydroxyl radical-promoted protein labeling efficiency.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
A series of phenol/aniline-derived probes have been synthesized, and their protein labeling behaviors in response to hydroxyl radical have been evaluated in test tubes. Among them, 2~3 probes showing high labeling efficiency and high selectivity to hydroxyl radical over other ROS were selected for live-cell experiments in the next stage. This progress is almost consistent with the research plan in the proposal.
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| Strategy for Future Research Activity |
This project is going to the second stage for the study of hydroxyl radical in neurodegenerative disease models, ranging from cultured cells to live mouse brain. The hydroxyl radical conditional protein labeling can be first evaluated under SDS-PAGE and western blot analysis. Confocal fluorescence imaging will be applied to identify the production sites of hydroxyl radical in the living systems. Furthermore, the labeled protein, which are associated with hydroxyl radical production, oxidation, and/or the downstream pathological pathways, can be profiled following a conditional proteomics workflow. The probe structures may require further optimization for its application in the complex living systems.
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| Causes of Carryover |
Due to the COVID-19 situation, order and delivery of some oversea products are canceled or delayed. Also, the domestic and international academic conferences were mostly held on-line, without travelling and accommodation fees to be paid. Given the COVID-19 situation is getting better, the unused amounts could be applied in the next fiscal year for the purchase of oversea products and on-site attending academic onferences.
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