{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Cabantous S"],"funding":["NIGMS NIH HHS"],"pagination":["2854"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC3790201"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["3"],"pubmed_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."],"journal":["Scientific reports"],"pubmed_title":["A new protein-protein interaction sensor based on tripartite split-GFP association."],"pmcid":["PMC3790201"],"funding_grant_id":["U54 GM074946","P01 GM098177","1 U54GM074946-01"],"pubmed_authors":["Pedelacq JD","Terwilliger TC","Koraichi F","Chaudhary A","Cabantous S","Ganguly K","Favre G","Nguyen HB","Waldo GS","Lockard MA"],"additional_accession":[]},"is_claimable":false,"name":"A new protein-protein interaction sensor based on tripartite split-GFP association.","description":"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.","dates":{"release":"2013-01-01T00:00:00Z","publication":"2013 Oct","modification":"2025-04-20T02:25:31.633Z","creation":"2019-03-27T01:16:51Z"},"accession":"S-EPMC3790201","cross_references":{"pubmed":["24092409"],"doi":["10.1038/srep02854"]}}