“CF-PPiD”, a new protein-protein interaction (PPI) screening technology using human protein bead arrays, was reported in an academic paper.
A new protein-protein interaction (PPI) screening technology “CF-PPiD“, which applies “proximity-dependent biotin labeling technology” to the “20K Human Protein Bead Array” containing 20,000 human-derived proteins synthesized by the wheat cell-free synthesis system, has been reported as following academic paper.
CF-PPiD technology based on cell-free protein array and proximity biotinylation enzyme for in vitro direct interactome analysis
Shusei Sugiyama, Kohdai Yamada, Miwako Denda, Satoshi Yamanaka, Satoshi Ozawa, Ryo Morishita & Tatsuya Sawasaki
Scientific Reports volume 12, Article number: 10592 (2022)
Protein–protein interaction (PPI) analysis is a key process to understand protein functions. Recently, we constructed a human protein array (20 K human protein beads array) consisting of 19,712 recombinant human proteins produced by a wheat cell-free protein production system. Here, we developed a cell-free protein array technology for proximity biotinylation-based PPI identification (CF-PPiD). The proximity biotinylation enzyme AirID-fused TP53 and -IκBα proteins each biotinylated specific interacting proteins on a 1536-well magnetic plate. In addition, AirID-fused cereblon was shown to have drug-inducible PPIs using CF-PPiD. Using the human protein beads array with AirID-IκBα, 132 proteins were biotinylated, and then selected clones showed these biological interactions in cells. Although ZBTB9 was not immunoprecipitated, it was highly biotinylated by AirID-IκBα, suggesting that this system detected weak interactions. These results indicated that CF-PPiD is useful for the biochemical identification of directly interacting proteins.