Abstract
Protein functions rely on their ability to engage in specific protein–protein interactions and form complexes that are dynamically regulated by stimuli. Bioluminescence resonance energy transfer (BRET) is a highly sensitive technique, which allows monitoring of interaction between two proteins: one tagged with the luminescent donor Renilla luciferase, the other with a fluorescent acceptor such as YFP. We adapted this method to single-cell imaging. To this aim, we tag proteins of interest, transfect cells with these fusions, and use the high-sensitivity microscopy, combined with electron multiplying cooled charge-coupled device (EMCCD) cameras and improved bioluminescence probes. We thus achieve rapid acquisition of high-resolution BRET images and study the localization and dynamics of protein–protein interactions in individual live cells.
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Perroy, J. (2010). Subcellular Dynamic Imaging of Protein–Protein Interactions in Live Cells by Bioluminescence Resonance Energy Transfer. In: Papkovsky, D. (eds) Live Cell Imaging. Methods in Molecular Biology, vol 591. Humana Press. https://doi.org/10.1007/978-1-60761-404-3_19
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DOI: https://doi.org/10.1007/978-1-60761-404-3_19
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Online ISBN: 978-1-60761-404-3
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