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Updated: Jan 8, 2026

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

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Published on: November 28, 2017

氨酸激活的裂变性可以促进交叉链接质谱学,用于对内源性蛋白质-蛋白质相互作用的全球分析

Fenglong Jiao¹, Merav Braitbard², Clinton Yu¹

¹Department of Physiology and Biophysics, University of California, Irvine, CA, USA.

Nature communications

|December 12, 2025

概括

这项研究引入了一种新的基于氨酸的可切割交叉链接质谱 (XL-MS) 平台,以增强蛋白质-蛋白质相互作用映射. 新方法显著扩大了蛋白质组覆盖范围,并揭示了对细胞网络结构的洞察力.

科学领域:

化学生物学化学生物学
蛋白质组学是指蛋白质组学.
结构生物学结构生物学

背景情况:

交联质谱 (XL-MS) 对于研究蛋白质-蛋白质相互作用 (PPI) 和蛋白质复合结构至关重要.
目前的XL-MS方法在蛋白质组覆盖范围上有局限性,这阻碍了全面的相互作用组分析.

研究的目的:

开发一种基于氨酸的创新可切割的XL-MS平台,以扩大全球PPI分析.
通过氧化诱导的氨酸可裂性来实现交叉链接的明确识别.

主要方法:

使用的非可切割的异基生物功能氨酸-氨酸 (K-C) 交叉链接器.
应用一种基于氨酸的新型可切割的XL-MS策略,对完整的细胞进行全蛋白质组分析.
采用了三种不同的异性双功能交叉链接器来证明其广泛适用性.

主要成果:

在2007年蛋白质中确定了25401个独特的联系,大大增强了蛋白质-蛋白质相互作用地图.
增加了人类互动组的相互连接性.
集成的XL-MS数据与AlphaFold预测和整合建模,以表征细胞网络和蛋白质复合体.

结论:

开发的基于氨酸的可切割XL-MS平台显著提升了PPI分析和蛋白质覆盖范围.
该策略提供了对原生蛋白质复合体的结构洞察,包括SERBP1-核糖体和eEF1A1-eEF1B.
这种方法为开发新的交联试剂和在化学蛋白质组学中表征氨酸修饰提供了基础.

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相关概念视频

Protein Networks

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.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...