Defining dynamic protein interactions using SILAC-based quantitative mass spectrometry.
Defining dynamic protein interactions using SILAC-based quantitative mass spectrometry.
复制标题
使用基于 SILAC 的定量质谱法定义动态蛋白质相互作用。
DOI：
10.1007/978-1-4939-1142-4_14
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Huang,Lan
中科院分区：
文献类型：
--
作者：
Wang,Xiaorong;Huang,Lan
蛋白质-蛋白质相互作用对于活细胞中的各种生理过程是必不可少的。蛋白质相互作用的完整表征对于我们理解它们在蛋白质功能调节中的作用至关重要。亲和纯化-质谱联用技术（AP-MS）已成为系统研究蛋白质相互作用的最有效方法之一。结合定量质谱，特异性相互作用蛋白质可以有效地从非特异性背景蛋白质区分。基于相互作用亲和力和动力学，蛋白质相互作用可以分为不同的类别，如稳定和动态相互作用。标准的生物化学方法在捕获和鉴定稳定的蛋白质相互作用方面是有效的，但不足以鉴定动态相互作用物。在这一章中，我们描述了集成策略，使识别蛋白质复合物的动态相互作用，将新的样品制备方法与SILAC为基础的定量。
Protein–protein interactions are essential to various physiological processes in living cells. A full characterization of protein interactions is critical to our understanding of their roles in the regulation of protein functions. Affinity purification coupled with mass spectrometry (AP-MS) has become one of the most effective approaches to systematically study protein–protein interactions. In combination with quantitative mass spectrometry, specific interacting proteins can be efficiently distinguished from nonspecific background proteins. Based on interaction affinity and kinetics, protein interactions can be classified into different categories such as stable and dynamic interactions. Standard biochemical methods are effective in capturing and identifying stable protein interactions but are not sufficient enough to identify dynamic interactors. In this chapter, we describe integrated strategies to allow the identification of dynamic interactors of protein complexes by incorporating new sample preparation methods with SILAC-based quantitation.