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发现蛋白质与蛋白质之间的相互作用,使用共分化质谱与无标签量化

Mopelola O Akinlaja^1,2, R Greg Stacey², Queenie W T Chan²

¹Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada.

Methods in molecular biology (Clifton, N.J.)

|July 14, 2023

概括

这项研究提出了一种新的无标签同分离质谱法 (CF-MS) 协议,用于识别蛋白质与蛋白质之间的相互作用,而无需改变样本. 该方法使用尺寸排除色谱和生物信息学管道来绘制蛋白质复合物的地图.

科学领域:

分子生物学分子生物学
生物化学生化学
蛋白质组学是指蛋白质组学.

背景情况:

蛋白质相互作用是分子生物学的基础.
同分离质谱法 (CF-MS) 是研究这些相互作用的关键技术.
现有的CF-MS协议通常需要样本标签,限制其适用性.

研究的目的:

描述一个新的,无标签的同分离质谱法 (CF-MS) 协议.
提供详细的方法来识别无需样本修改的蛋白质-蛋白质相互作用.
引入PrInCE生物信息管道用于蛋白质复合体预测.

主要方法:

细胞或组织在非变性条件下被溶解.
完整的蛋白质复合体是通过大小排除色谱 (SEC) 分离的.
每个分量的蛋白质都通过质谱分析,并使用生物信息学管道 (PrInCE) 进行分析.

主要成果:

该协议允许无标签检测蛋白质与蛋白质相互作用.
它允许识别和量化数千种蛋白质相互作用.
普林斯管道准确地从CF-MS数据中预测蛋白质复合体.

结论:

无标签的CF-MS是一种可行且强大的方法,用于蛋白质组范围内的相互作用研究.
该协议为现有的标记方法提供了有价值的替代方案.
结合CF-MS和生物信息学工具,提高了蛋白质复合物的发现.

关键词:

协同分成的分成法.没有标签的量化.质谱测量质量谱测量蛋白质蛋白质相互作用尺寸排除色谱学尺寸排除色谱学

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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,...