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

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, E. coli, 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 in vitro and in vivo, monitoring the biological process, and controlling the activity of the enzyme of interest.

蛋白片段互补分析技术研究进展

基于报告蛋白功能重建的蛋白片段互补分析(protein fragment complementation assay,PCA)技术是研究体内蛋白质相互作用重要方法,可检测和分析蛋白相互作用及其时空变化和调节因素。目前已开发出多种不同报告蛋白的PCA系统,具有灵敏度高、信噪比高、可定量和高通量化、适用范围广等特点。主要对PCA的原理、不同类型及其应用领域等方面进行阐述,并展望其发展前景。