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

已开发了多种方法来检测、分析和定量蛋白质相互作用，包括SPR、ITC、共沉淀法、亲和层析、超速离心法、核磁共振、质谱仪和ELISA。虽然这些方法被广泛用于识别蛋白质的功能，但当涉及到生物途径的复杂、动态机制时，它们很快就达到了极限。相比之下，基于细胞的PPI分析适用于更广泛的应用。但是，这些检测方法都有不同的局限性，如成像对比度低、使用固定细胞、成本高、需要进行大量的阶梯式实验。在这项研究中，我们建立了PPI检测系统，该系统在活细胞中观察在生理环境下与结合伙伴结合的位点特异性标记的靶蛋白。

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.