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一个照片的交叉链接器用于识别蛋白质与蛋白质之间的相互作用

Xingyu Chen^1,2, Cristina Jiménez López³, André Nadler³

¹Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.

Chembiochem : a European journal of chemical biology

|November 21, 2024

概括

研究人员开发了一种新的光响应交叉连接器,用于使用交叉连接质谱法 (XL-MS) 改进蛋白质-蛋白质相互作用 (PPI) 研究. 这种新工具为细胞内PPI分析提供了增强的细胞透性和时间控制.

科学领域:

生物化学生化学
化学生物学化学生物学
蛋白质组学是指蛋白质组学.

背景情况:

交联质谱 (XL-MS) 对于研究蛋白质与蛋白质相互作用 (PPI) 是至关重要的.
目前的交叉连接器面临着膜透性和副作用的限制,阻碍了细胞内PPI分析.
需要具有增强稳定性,疏水性和时间控制的交叉连接器.

研究的目的:

为XL-MS.合成和验证一种新的光响应交叉连接器.
克服现有的交叉连接器在细胞内PPI研究的局限性.
为了实现精确的,UV触发蛋白质交联与时间分辨率.

主要方法:

一种新型交叉连接剂的合成,其中包括o-nitrobenzyl ester (o-NBE) 的光响应组.
紫外线触发的交叉连接器的激活,形成化物产品.
阿尔德与相邻的氨基基团的反应以及随后的还原性氨基化,形成稳定的二次氨基链接.
使用重组模型蛋白和XL-MS的原则证明实验.
在完整的HeLa细胞中评估细胞透性.

主要成果:

该o-NBE交叉连接器表现出增强的稳定性和疏水性质.
紫外线成功触发了与时间控制的交联反应.
交叉连接器可靠地通过XL-MS在重组模型蛋白中检测到PPI.
该o-NBE交联器显示能够进入完整的HeLa细胞.

结论:

新的o-NBE交叉连接器为研究具有增强功能的PPI提供了强大的工具.
它的光响应性和细胞透性为细胞内PPI分析提供了显著的优势.
这种交叉连接器有可能促进细胞环境和药物发现的研究.

关键词:

交叉连接器交叉连接器质谱测量质量谱测量蛋白质与蛋白质的相互作用蛋白质组学是指蛋白质组学.没有被关在子里.

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Protein-protein Interfaces

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Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a...

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Protein Networks

An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
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