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A New Protein-Protein Interaction Sensor Based on Tripartite Split-GFP Association

Abstract : Monitoring protein-protein interactions in living cells is key to unraveling their roles in numerous cellular processes and various diseases. Previously described split-GFP based sensors suffer from poor folding and/ or self-assembly background fluorescence. Here, we have engineered a micro-tagging system to monitor protein-protein interactions in vivo and in vitro. The assay is based on tripartite association between two twenty amino-acids long GFP tags, GFP10 and GFP11, fused to interacting protein partners, and the complementary GFP1-9 detector. When proteins interact, GFP10 and GFP11 self-associate with GFP1-9 to reconstitute a functional GFP. Using coiled-coils and FRB/FKBP12 model systems we characterize the sensor in vitro and in Escherichia coli. We extend the studies to mammalian cells and examine the FK-506 inhibition of the rapamycin-induced association of FRB/FKBP12. The small size of these tags and their minimal effect on fusion protein behavior and solubility should enable new experiments for monitoring protein-protein association by fluorescence.
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Contributor : Jean-Denis Pedelacq <>
Submitted on : Tuesday, November 24, 2020 - 3:23:19 PM
Last modification on : Tuesday, January 19, 2021 - 11:06:43 AM
Long-term archiving on: : Thursday, February 25, 2021 - 8:23:24 PM


Scientific Report 2013.pdf
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Stéphanie Cabantous, Hau Nguyen, Jean-Denis Pedelacq, Faten Koraïchi, Anu Chaudhary, et al.. A New Protein-Protein Interaction Sensor Based on Tripartite Split-GFP Association. Scientific Reports, Nature Publishing Group, 2013, 3 (1), ⟨10.1038/srep02854⟩. ⟨hal-02378714⟩



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