2013 Fiscal Year Final Research Report
Development of bioluminescence methods for monitoring autophagy in living cells
| Project/Area Number |
24710259
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Chemical biology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
HATTORI Mitsuru 東京大学, 理学(系)研究科(研究院), 研究員 (20589858)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Keywords | ルシフェラーゼ / オートファジー |
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| Research Abstract |
We developed a new bioluminescence monitoring system for detecting autophagy in living cells. We have an optical technique for monitoring protein-protein interactions in living cells based on complementation of split fragments of luciferase. By connecting proteins that composed autophagosome membrane with the luciferase fragments, we were able to develop the system to detect the conformation of autophagosome as a model of autophagy.
These probes were induced to human cultured cells and measured the luminescence with serum starvation. Each pairs showed significant increasing of luminescence counts. We also arranged the probes for improving the luminescence intensity. By exchanging the luciferase to other type, the luminescence intensity was increased about 100 times than old types. Consequently, the method enables rapid and reliable approach for the monitoring of autophagy in cells and living subjects.
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[Journal Article] Bioluminescent Probes to Analyze Ligand- induced Phosphatidylinositol 3,4,5- trisphosphate Production with Split Luciferase Complementation Anal2013
Author(s)
Yang, L. Z., Nasu, Y., Hattori, M., Yoshimura, H., Kanno, A. and Ozawa, T
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Journal Title
Chem
Volume: 85
Pages: 11352-11359
DOI
Peer Reviewed
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[Journal Article] ROS stress resets circadian clocks to coordinate pro- survival signals2013
Author(s)
Tamaru, T., Hattori, M., Ninomiya, Y., Kawamura, G., Vares, G., Honda, K., Mishra, D. P., Wang, B., Benjamin, I., Sassone- Corsi, P., Ozawa, T. and Takamatsu, K
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Journal Title
PLoS ONE
Volume: 8
Pages: e82006
DOI
Peer Reviewed
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[Journal Article] Analysis of temporal patterns of GPCR-β- arrestin interactions using split luciferase- fragment complementation Mol2013
Author(s)
Hattori, M., Tanaka, M., Takakura, H., Aoki, K., Miura, K., Anzai, T. and Ozawa, T
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Journal Title
BioSyst
Volume: 9
Pages: 957-964
DOI
Peer Reviewed
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