Development of a novel screening system for PPI-targeted drugs using cutinase fusion strategy

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A wide variety of methods have been developed to detect, analyze, and quantify protein interactions, including SPR, ITC, co-precipitation, affinity chromatography, ultracentrifugation, NMR, mass spectrometry, and ELISAs in vitro. While these methods are widely used to identify protein function, they quickly reach their limits when it comes to addressing the intricate, dynamic mechanics of a biological pathway. In contrast, cell-based PPIs assays are appropriate for wider applications. However, these assays have different limitations such as the low-contrast imaging, using fixed-cells, high-cost and a lot of step experiments. In this study, we have established the PPI assay system which observes the site-specifically labeled target proteins with a binding partner under physiological context in live cells.