激酶催化的生物素化作用。
Kinase-Catalyzed Biotinylation.
作者信息
Senevirathne Chamara, Green Keith D, Pflum Mary Kay H
机构信息
Department of Chemistry, Wayne State University, Detroit, MI, 48202.
出版信息
Curr Protoc Chem Biol. 2012;4(1):83-100. doi: 10.1002/9780470559277.ch110228.
Abstract
Kinase-catalyzed protein phosphorylation plays an essential role in a variety of biological processes. Methods to detect phosphoproteins and phosphopeptides in cellular mixtures will aid in cell biological and signaling research. Our laboratory recently discovered the utility of γ-modified ATP analogues as tools for studying phosphorylation. Specifically, ATP-biotin can be used for labeling and visualizing phosphoproteins from cell lysates. Because the biotin tag is suitable for protein detection, the biotinylation reaction can be applied to multiple phosphoproteomics applications. Herein we report a general protocol for labeling phosphopeptides and phosphoproteins in biological samples using kinase-catalyzed biotinylation.
摘要
激酶催化的蛋白质磷酸化在多种生物学过程中起着至关重要的作用。检测细胞混合物中磷酸化蛋白和磷酸化肽的方法将有助于细胞生物学和信号转导研究。我们实验室最近发现γ-修饰的ATP类似物作为研究磷酸化的工具的效用。具体而言，ATP-生物素可用于标记和可视化细胞裂解物中的磷酸化蛋白。由于生物素标签适用于蛋白质检测，生物素化反应可应用于多种磷酸化蛋白质组学应用。在此，我们报告了一种使用激酶催化生物素化标记生物样品中磷酸化肽和磷酸化蛋白的通用方案。
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