AlphaScreen Technology for High Throughput Screenin(RMI)

描述(申请人提供)：14-3-3蛋白通过依赖磷酸化与靶蛋白结合，调节广泛的生理过程，包括细胞增殖、细胞周期进程、抑制凋亡程序和神经元功能。为了开发研究14-3-3在各种生物系统中的功能的研究工具，并有可能开发14-3-3靶向治疗策略，我们开发了一种基于AlphaScreen技术的高通量分析方法，用于分离能够破坏14-3-3/客户蛋白相互作用的小分子。我们已经优化和微型化了适合机器人手术的384孔板格式的检测方法。这项建议的目标是在分子文库筛选中心网络中利用这种测试来分离可能干扰14-3-3与其客户蛋白相互作用的小分子化合物。

DESCRIPTION (provided by applicant): Through phosphorylation-dependent binding to target proteins, 14-3-3 proteins regulate a wide range of physiological processes, including cell proliferation, cell cycle progression, suppression of apoptotic program, and neuronal functions. In order to generate research tools for study the functions of 14-3-3 in various biological systems and potentially to develop 14-3-3-targeted therapeutic strategies, we have developed an AlphaScreen technology based high throughput assay for isolation of small molecules that can disrupt the 14-3-3/client protein interactions. We have optimized and miniaturized the assay for adaptation in a 384-well plate format for robotic operations. The goal of this proposal is to utilize this assay in the Molecular Libraries Screening Centers Network for isolation of small molecule compounds that can interfere with the interaction of 14-3-3 with its client proteins.