TITLE:
Development of Split-Protein Systems: From Binary to Ternary System
AUTHORS:
Shengyi Shen
KEYWORDS:
Split-Protein Reassembly, Protein Fragment Complementation, Chemical In-ducer of Dimerization (CID), Protein-Protein Interaction (PPI)
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.12 No.3,
March
26,
2021
ABSTRACT: 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.