Development of Split-Protein Systems: From Binary to Ternary System  

作　　者：Shengyi Shen Shengyi Shen(Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA)

机构地区：[1]Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA

出　　处：《Advances in Bioscience and Biotechnology》2021年第3期78-94,共17页生命科学与技术进展（英文）

摘　　要：Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the methods to catalogue these macromolecules by detecting their interactions, modifications, and cellular locations. It will be helpful for scientists to compare the difference between a diseased cellular state and its normal state and to find the potential therapy treatment to intervene this status. One technology called split-protein reassembly or protein fragment complementation has been developed in the last two decades. This technology makes use of appropriate fragmentation of some protein reporters and the refolding of these reports could be detected by their function to confirm the interaction of interest. This system has been set up in cell-free systems, </span><i><span style="font-family:Verdana;">E.</span></i></span><i><span style="font-family:""> </span></i><i><span style="font-family:Verdana;">coli</span></i><span style="font-family:""><span style="font-family:Verdana;">, yeast, mammalian cells, plants and live animals. Herein, I present the development in fluorescence- and bioluminescence-based split-protein biosensors in both binary and ternary systems. In addition, some people developed the split-protein system by combining it with chemical inducer of dimerization strategy (CID). This has been applied for identifying the enzyme inhibitors and regulating the activity of protein kinases and phosphatases. With effort from many laboratories from the world, a variety of split-protein systems have been developed for studying the PPI </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> and </span><i><span style="font-family:Verdana;">in vivo</span></i><span style="font-family:Verdana;">, monitoring the biological process, and controlling the activity of the enzyme of interest.Tens of thousands of protein-protein interactions (PPIs) have been found in human cells and many of these macromolecular partnerships could determine the cell growth and death. Thus there is a need to develop the methods to catalogue these macromolecules by detecting their interactions, modifications, and cellular locations. It will be helpful for scientists to compare the difference between a diseased cellular state and its normal state and to find the potential therapy treatment to intervene this status. One technology called split-protein reassembly or protein fragment complementation has been developed in the last two decades. This technology makes use of appropriate fragmentation of some protein reporters and the refolding of these reports could be detected by their function to confirm the interaction of interest. This system has been set up in cell-free systems, </span><i><span style="font-family:Verdana;">E.</span></i></span><i><span style="font-family:""> </span></i><i><span style="font-family:Verdana;">coli</span></i><span style="font-family:""><span style="font-family:Verdana;">, yeast, mammalian cells, plants and live animals. Herein, I present the development in fluorescence- and bioluminescence-based split-protein biosensors in both binary and ternary systems. In addition, some people developed the split-protein system by combining it with chemical inducer of dimerization strategy (CID). This has been applied for identifying the enzyme inhibitors and regulating the activity of protein kinases and phosphatases. With effort from many laboratories from the world, a variety of split-protein systems have been developed for studying the PPI </span><i><span style="font-family:Verdana;">in vitro</span></i><span style="font-family:Verdana;"> and </span><i><span style="font-family:Verdana;">in vivo</span></i><span style="font-family:Verdana;">, monitoring the biological process, and controlling the activity of the enzyme of interest.

关 键 词：Split-Protein Reassembly Protein Fragment Complementation Chemical In-ducer of Dimerization (CID) Protein-Protein Interaction (PPI) 

分 类 号：R73[医药卫生—肿瘤]