Abstract
Post-translational modification, including phosphorylation and ubiquitination, plays a pivotal role in intracellular signal transduction. During the past few decades, post-translational modification has mainly been analyzed by biochemical methods such as immunoblotting. However, with the advent of green fluorescent protein (GFP) and its color variants, fluorescence imaging has provided spatial and temporal information on protein localization in living cells. In addition, Förster (or fluorescence) resonance energy transfer (FRET)-based biosensors enable us to visualize post-translational modifications, contributing to the progress in the research field of post-translational modification and intracellular signaling. Here, I review the principles of FRET, the design of the FRET biosensor, and its applications. Finally, I discuss the future perspectives of FRET biosensors for post-translational modification research.
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Acknowledgments
I thank K. Takakura, N. Nishimoto, Y. Inaoka, K. Hirano, and A. Kawagishi for their technical assistance. I also thank the members of the Matsuda Laboratory for their helpful discussions. K.A. was supported by the Platform for Dynamic Approaches to Living System from the Ministry of Education, Culture, Sports, and Science, Japan, and by a Grant-in-Aid for Scientific Research on Innovative Areas (25117715). The author declares no conflict of interest.
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Aoki, K. (2015). Visualization of Intracellular Signaling with Fluorescence Resonance Energy Transfer-Based Biosensors. In: Inoue, Ji., Takekawa, M. (eds) Protein Modifications in Pathogenic Dysregulation of Signaling. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55561-2_3
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DOI: https://doi.org/10.1007/978-4-431-55561-2_3
Publisher Name: Springer, Tokyo
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