通过近距离增强的生物对角联结对特定蛋白质糖形的活细胞标记
Peter V Robinson1, Gabriela de Almeida-Escobedo1, Amber E de Groot1
1Departments of Chemistry and Molecular and Cell Biology and the Howard Hughes Medical Institute, University of California , Berkeley, California 94720, United States.
Journal of the American Chemical Society
|August 18, 2015
概括
我们开发了循环octyne-aptamer结合物来检测活细胞中的特定蛋白质糖化. 这种新的方法可以使用各种检测技术精确分析糖化表位.
科学领域:
- 生物化学
- 分子生物学
- 化学生物学
背景情况:
- 在特定蛋白质上检测翻译后的修饰,特别是糖化,是具有挑战性的.
- 现有的方法往往缺乏蛋白质背景中的糖化表位的特异性.
研究的目的:
- 开发一种用于检测蛋白质特异性糖化的一种新方法.
- 创建可以选择性地向和检测活细胞上的糖化表位体的试剂.
主要方法:
- 用于选择性结合到标记为azidosugar的甘氨酸的循环octyne-aptamer联合体.
- 应用该方法以准特定的细胞表面糖蛋白.
- 使用质谱测量,西方抹杀和流细胞测量进行了验证检测.
主要成果:
- 在目标蛋白质上证明了对胺结合物的选择性结合.
- 对两种不同的细胞表面糖蛋白进行了成功的特征化.
- 展示了试剂在检测蛋白质形中的实用性.
结论:
- 循环octyne-aptamer结合物为检测特定蛋白质的糖化提供了一个强大的平台.
- 这项技术有助于研究糖化在细胞过程中的作用.
- 这种方法可以适应研究各种细胞表面标和糖化模式.
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