Dynamic Imaging of Small Molecule Induced Protein-Protein Interactions in Living Cells with a Fluorophore Phase Transition Based Approach.
Dynamic Imaging of Small Molecule Induced Protein-Protein Interactions in Living Cells with a Fluorophore Phase Transition Based Approach.
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基于荧光团相变的活细胞中小分子诱导的蛋白质-蛋白质相互作用的动态成像方法。
DOI：
10.1021/acs.analchem.8b03476
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发表时间：
2018-12-18
影响因子：
7.4
通讯作者：
Shu X
中科院分区：
文献类型：
--
作者：
Chung CI;Zhang Q;Shu X
Protein-protein interactions (PPIs) mediate signal transduction in cells. Small molecules that regulate PPIs are important tools for biology and biomedicine. Dynamic imaging of small molecule-induced PPIs characterizes and verifies these molecules in living cells. It is thus important to develop cellular assays for dynamic visualization of small molecule-induced protein-protein association and dissociation in living cells. Here we have applied fluorophore phase transition-based principle and designed a PPI assay named SPPIER (separation of phases-based protein interaction reporter). SPPIER utilizes the green fluorescent protein (GFP) and is thus genetically encoded. Upon small molecule-induced PPI, SPPIER rapidly forms highly fluorescent GFP droplets in living cells. SPPIER detects immunomodulatory drugs (IMiDs)-induced PPI between cereblon and the transcription factor Ikaros. It also detects IMiDs analog (e.g. CC-885)-induced PPI between cereblon and GSPT1. Furthermore, SPPIER can visualize bifunctional molecules (e.g. PROTAC)-induced PPI between an E3 ubiqutin ligase and a target protein. Lastly, SPPIER can be modified to image small molecule-induced protein-protein dissociation, such as nutlin-induced dissociation between HDM2 and p53. The intensive brightness and rapid kinetics of SPPIER enable robust and dynamic visualization of PPIs in living cells.
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DOI：
10.1126/science.aab1433
发表时间：
2015-06-19
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Winter GE;Buckley DL;Paulk J;Roberts JM;Souza A;Dhe-Paganon S;Bradner JE
通讯作者：
Bradner JE
影响因子：
56.9
作者：
Ito, Takumi;Ando, Hideki;Handa, Hiroshi
通讯作者：
Handa, Hiroshi
影响因子：
64.8
作者：
Li, Pilong;Banjade, Sudeep;Cheng, Hui-Chun;Kim, Soyeon;Chen, Baoyu;Guo, Liang;Llaguno, Marc;Hollingsworth, Javoris V.;King, David S.;Banani, Salman F.;Russo, Paul S.;Jiang, Qiu-Xing;Nixon, B. Tracy;Rosen, Michael K.
通讯作者：
Rosen, Michael K.
影响因子：
56.9
作者：
Vassilev, LT;Vu, BT;Liu, EA
通讯作者：
Liu, EA
DOI：
10.1126/science.1198841
发表时间：
2011-05-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Grigoryan G;Kim YH;Acharya R;Axelrod K;Jain RM;Willis L;Drndic M;Kikkawa JM;DeGrado WF
通讯作者：
DeGrado WF