使用Prod&PQ-seqq进行多重测量蛋白质-蛋白质相互作用和蛋白质丰富度在细胞条件的多重测量
bioRxiv : the preprint server for biology
|January 9, 2026
概括
Prod&PQ-seq是一种研究蛋白质与蛋白质相互作用 (PPI) 和蛋白质水平的新方法. 这种可扩展的框架量化了PPI和蛋白质丰度在各种条件下的变化.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 目前的蛋白质与蛋白质相互作用 (PPI) 分析方法缺乏可扩展性,并且仅限于少数条件或目标.
- 在不同的生物环境中有效研究PPI和蛋白质定量对于理解细胞机制至关重要.
研究的目的:
- 引入Prod&PQ-seq,这是一个用于多重检测和PPI和蛋白质量测量的新框架.
- 为了证明Prod&PQ-seq. 的定量性,可重复性,敏感性和特异性.
主要方法:
- Prod&PQ-seq使用交叉链接的细胞,抗体-寡核酸合物 (ab-oligos),以及用于PPI捕获的DNA校准器.
- 该框架允许近接ab-oligos的双向原始化,以检测和量化相互作用.
- 基准测试涉及复合复合物,定位和细胞混合实验.
主要成果:
- Prod&PQ-seq被验证为一种定量,可复制,敏感和特定的方法.
- 应用到多抑制复合物2 (PRC2) 显示,EZH2抑制和H3.3K27M表达减弱了PRC2-H3K27me3相互作用和PRC2内部的PPI.
- 对H3.1K27M和H3.3K27M变体观察到明显的PPI配置文件,影响了像H3K27ac-K27M和H3K27ac-EED这样的相互作用.
结论:
- 在Prod&PQ-seq中,可以检测PPI成分和强度的变化.
- 该框架促进了跨生物条件,小分子抑制和遗传干扰的蛋白质量化.
- Prod&PQ-seq提供了一个可扩展的解决方案,用于全面的PPI和蛋白质分析.
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