| Project/Area Number |
13554030
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
分離・精製・検出法
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| Research Institution | The University of Tokyo |
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Principal Investigator |
UMEZAWA Yoshio The University of tokyo, Graduate School of Science, Professor, 大学院・理学系研究科, 教授 (80011724)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Moritoshi The University of tokyo, Graduate School of Science, Assistant, 大学院・理学系研究科, 助手 (00323501)
OZAWA Takeaki The University of tokyo, Graduate School of Science, Lecturer, 大学院・理学系研究科, 講師 (40302806)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥14,300,000 (Direct Cost: ¥14,300,000)
Fiscal Year 2002: ¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 2001: ¥10,200,000 (Direct Cost: ¥10,200,000)
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| Keywords | fluorescent probes / protein / protein interaction / non-destructive analysis / second messengers / protein phosphorylation / ミトコンドリア / エストロゲン / GFP / 蛋白質間相互作用 / ルシフェラーゼ / 疎水場感受性蛍光試薬 / サイクリックGMP |
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| Research Abstract |
For nondestructive analysis of chemical processes in living cells, we have developed some general methods and new intracellular fluorescent probes for detecting 1) second messengers, cGMP, diacylglycerol and phosphatidylinositol-3,4,5-triphosphate, 2) protein phosphorylation, 3) protein confbrmational changes, 4) protein-protein interactions, and 5) protein localizations in live cells under a confocal laser microscope.
The approaches for the present probe developments are use of fluorescence resonance energy transfer for reporting binding of substrates (analytes) to molecular recognition domains in dual-fluorophore conjugated probe molecules, and use of protein splicing chemistry for detecting protein-protein interactions.
Key molecules and steps of cellular signaling pathways were visualized in relevant live cells using developed fluorescent probe molecules. These probes were found of general importance not only for fundamental biology studies, but also for assay and screening methods for chemicals that inhibit or facilitate cellular signaling pathways.
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