| Project/Area Number |
22655055
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemistry related to living body
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| Research Institution | Osaka University |
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Principal Investigator |
OHKANDA Junko 大阪大学, 産業科学研究所, 准教授 (50233052)
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| Project Period (FY) |
2010 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2010: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | 蛍光プローブ / 天然物 / リン酸化ペプチド / 14-3-3たんぱく質 / たんぱく質間相互作用 / フシコクシン / 抗がん活性 / 標的同定 / たんぱく質-たんぱく質相互作用 |
|---|
| Research Abstract |
14-3-3 proteins play a critical role in serine/ threonine kinase-dependent signaling pathways through protein-protein interactions with multiple phosphorylated ligands. A ligand-dependent 14-3-3 detection technique would facilitate elucidation of 14-3-3-related intracellular signaling networks. Here, we describe phosphopeptide-dependent fluorescent labeling of 14-3-3. using cell penetrating probes derived from a diterpene natural product, fusicoccin. In vitro evaluations demonstrated that these compounds site-specifically attached a fluorescent tag onto the protein surface as a result of ternary complex formation with the 14-3-3 protein and a phosphopeptide ligand. The BODIPY-attached probe labeled human endogenous 14-3-3 in cancer cells under hyper-phosphorylation conditions, proving that 14-3-3 is a primary target of the fusicoccins in mammalian cells. This cell-penetrating labeling agent would be a useful tool to explore the mechanism of antitumor activity associated with fusicoccin-related agents.
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