Abstract
There is a significant focus on detecting and assaying protein–protein interactions (PPIs) in biology and biotechnology. Protein-fragment complementation assay (PCA) is one of the most widely used methods to detect PPI by splitting the enzyme-coding or fluorescent protein-coding polypeptide, as well as Förster resonance energy transfer (FRET). Here, we describe a novel PPI assay FlimPIA (firefly luminescent intermediate-based protein–protein interaction assay) by a unique approach of splitting the two major catalytic steps (half reactions) of firefly luciferase (FLuc).
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Acknowledgment
We thank B. Branchini for providing inspiration for the project. This project was supported by SENTAN, JST, Japan, and partly by the “Leave a Nest” Microtech Nichion award.
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Ohmuro-Matsuyama, Y., Ueda, H. (2016). A Protein–Protein Interaction Assay FlimPIA Based on the Functional Complementation of Mutant Firefly Luciferases. In: Kim, S. (eds) Bioluminescence. Methods in Molecular Biology, vol 1461. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3813-1_10
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DOI: https://doi.org/10.1007/978-1-4939-3813-1_10
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