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通过使用MP3-seqq进行测序来大规模平行测量蛋白质-蛋白质相互作用

Alexandr Baryshev¹, Alyssa La Fleur², Benjamin Groves¹

¹Department of Electrical & Computer Engineering, University of Washington, Seattle, WA, USA.

Nature chemical biology

|August 27, 2024

概括

我们通过测序 (MP3-seq) 开发了大规模并行蛋白质-蛋白质相互作用,这是一个可扩展的酵母双混合方法. MP3-seq准确地测量了数千种蛋白质相互作用,有助于细胞和基因治疗的发展.

科学领域:

生物化学生化学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

蛋白与蛋白相互作用 (PPI) 对细胞功能至关重要.
工程化PPI对细胞和基因疗法有很大的前景.
需要准确和可扩展的方法来测量PPI.

研究的目的:

通过测序 (MP3-seq) 引入大规模并行PPI测量,这是一种新的酵母两混合 (Y2H) 方法.
展示MP3-seq的可扩展性和定量准确性,用于高通量PPI分析.
应用MP3-seq来描述设计的蛋白质相互作用和线圈-线圈特异性.

主要方法:

开发了MP3-seq,一个Y2H系统,利用与蛋白质对相关的DNA条形码.
通过测序条形码丰富量化量化相互作用强度.
应用MP3-seq来研究异构体相互作用和卷轴-卷轴特异性.
使用AlphaFold-Multimer (AF-M) 进行结构预测,并训练模型以能量计算.

主要成果:

MP3-seq具有高度定量性,可扩展到超过10万次交互.
在设计异构聚合物家族中的特征性相互作用.
在卷轴-卷轴相互作用中研究的特异性决定因素.
AF-M模型显示了预先选的潜力,但实验验证对于定量排名至关重要.

结论:

MP3-seq提供了一个强大的,可扩展的工具来测量PPI.
这种方法有助于研究用于治疗应用的工程相互作用.
计算模型可以补充实验数据,但不能取代定量相互作用测量.

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