Identification of protein-protein interactions by surface plasmon resonance followed by mass spectrometry.
Identification of protein-protein interactions by surface plasmon resonance followed by mass spectrometry.
复制标题
通过表面等离子共振和质谱法鉴定蛋白质-蛋白质相互作用。
DOI：
--
复制
发表时间：
2011
影响因子：
--
通讯作者：
P. Svenningsson
中科院分区：
文献类型：
--
作者：
A. Madeira;E. Vikeved;A. Nilsson;B. Sjögren;P. Andrén;P. Svenningsson
阐明蛋白质网络的功能和意义有助于理解许多病理过程和确定新的治疗靶点。本单元描述了一种通过耦合表面等离子体共振到质谱法来发现蛋白质-蛋白质相互作用的方法。简单地说，将蛋白质以共价结合到传感器芯片上，然后通过微流体系统将传感器芯片暴露在大脑提取液中。这样可以实时监测固定配体和提取物之间的相互作用。然后从提取物中回收相互作用的蛋白质，胰蛋白酶化，并使用质谱法鉴定。利用Mascot软件对序列数据库进行检索。为了排除非特异性相互作用，使用空白传感器芯片和/或随机肽进行对照实验。本文提出的方案不需要对蛋白质进行特定的标记或修饰，可以在4天内完成。
Elucidation of the function and meaning of the protein networks can be useful in the understanding of many pathological processes and the identification of new therapeutic targets. This unit describes an approach to discover protein-protein interactions by coupling surface plasmon resonance to mass spectrometry. Briefly, a protein is covalently bound to a sensor chip, which is then exposed to brain extracts injected over the surface via a microfluidic system. This allows the monitoring in real-time of the interactions between the immobilized ligand and the extracts. Interacting proteins from the extracts are then recovered, trypsinized, and identified using mass spectrometry. The data obtained are searched against a sequence database using the Mascot software. To exclude nonspecific interactors, control experiments using blank sensor chips, and/or randomized peptides, are performed. The protocol presented here does not require specific labeling or modification of proteins and can be performed in <4 days.
影响因子：
4.4
作者：
Nilsson, Anna;Skold, Karl;Andren, Per E.
通讯作者：
Andren, Per E.