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将SICLOPPS查与下一代测序集成在一起,以发现蛋白质-蛋白质相互作用的循环调节器

Juan Antonio Sarmiento Muro¹, Ali Tavassoli¹

¹Department of Chemistry, University of Southampton, Southampton, United Kingdom.

Methods in enzymology

|November 20, 2025

概括

这项研究引入了一种新的循环发现方法,该方法使用SICLOPPS (Split Intein Circular Ligation of Peptides and Proteins) 与下一代测序 (NGS) 相结合. 这种方法有效地识别了细胞内的蛋白质-蛋白质相互作用抑制剂.

科学领域:

分子生物学分子生物学
生物化学生化学
药物发现药物发现药物发现

背景情况:

蛋白与蛋白相互作用 (PPI) 对于细胞功能至关重要,但由于其复杂的接口,治疗目标具有挑战性.
循环正在成为有效的PPI抑制剂,提供刚性,亲和力和细胞透性.

研究的目的:

开发一种高通量方法,用于发现活细胞内抑制PPI的循环.
将和蛋白质循环结合 (SICLOPPS) 与下一代测序 (NGS) 集成,以实现高效的图书馆选和命中识别.

主要方法:

利用一个反向双混合系统 (RTHS) 与SICLOPPS生成的循环库进行功能选择.
采用代生物扫描与NGS相结合,用于定量分析和对所需序列的丰富.
开发了一个实时监测图书馆演变和过假阳性的管道.

主要成果:

展示了一个可扩展和可重复的工作流程,用于大规模发现循环类PPI抑制剂.
通过代选择和NGS量化成功识别了丰富的序.
展示了追踪库动态和在多个选择轮中改进命中识别的能力.

结论:

集成的SICLOPPS-NGS管道为发现细胞内PPI抑制剂提供了一个强大的平台.
这种方法显著扩大了调节蛋白质-蛋白质相互作用的工具包,在各种生物环境中具有广泛的适用性.
提供了一个强大的战略,用于识别针对PPI的新型治疗线索.

关键词:

生物制造是生物制造的一种.循环类酸是一种循环类酸.高通量选的高通量选这是下一代测序.类抑制剂类抑制剂蛋白质蛋白质相互作用一个叫做SICLOPPS的企业.

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