Abstract
We have established a method of video-rate bioluminescence imaging to visualize exocytotic protein secretion from a single living cell using Gaussia luciferase (GLase) as a reporter protein. The luminescence signals produced by the enzymatic reaction of secreted GLase (luciferase) and coelenterazine (luciferin) are detected with an electron-multiplying charge-coupled device camera. An exocytotic event of protein secretion can be visualized using the protein fused to GLase with a time resolution of 30–500 ms. Signal analyses of the bioluminescence video images reveal a number of exocytotic sites, the frequency of exocytotic events, and the amount of secreted protein on a whole cell. Furthermore, the method can distinguish between secreted and cell surface-bound proteins. Our method is a direct approach to investigate the secretion and localization of proteins on the whole surface of living cells.
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Acknowledgement
This work was supported in part by a Grant-in-Aid for Scientific Research (C) from the Japan Society for the Promotion of Science and by Strategic Research AGU-Platform Formation (2008–2012) to TS.
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Suzuki, T., Inouye, S. (2014). Video-Rate Bioluminescence Imaging of Protein Secretion from a Living Cell. In: Badr, C. (eds) Bioluminescent Imaging. Methods in Molecular Biology, vol 1098. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-718-1_6
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DOI: https://doi.org/10.1007/978-1-62703-718-1_6
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-717-4
Online ISBN: 978-1-62703-718-1
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