向全蛋白质组的照片交叉链接取得进展,以使生物体中蛋白质结构和网络的残留水平可视化
Anneliese M Faustino1, Piyoosh Sharma1, Edgar Manriquez-Sandoval2
1Department of Chemistry, Johns Hopkins University, Baltimore, Maryland 21218, United States.
Analytical chemistry
|June 21, 2023
概括
新的光交叉连接器使得细胞内的蛋白相互作用的全蛋白体研究成为可能. 这些工具以残留级分辨率识别蛋白质接触,推动细胞生物学研究.
科学领域:
- 结构生物学是结构生物学.
- 细胞生物学 细胞生物学
- 化学生物学是化学生物学.
背景情况:
- 交联质谱法 (XL-MS) 通过残留级分辨率识别蛋白质-蛋白质相互作用.
- 现有的MS可切割的交叉链接器可以促进体内蛋白质接触的识别.
- 照片交叉链接器提供高反应性和时间分辨率,但很难识别.
研究的目的:
- 为全蛋白质组研究合成和应用新型异构生物功能光交叉链接剂.
- 为了克服与照片交叉链接相关的识别挑战.
- 为了证明在纤维质中照片交叉链接的可行性.
主要方法:
- 合成含有迪亚齐林和N-基-苏奇尼米迪尔碳酸盐基的异构功能光交叉链接剂.
- 使用MS-cleavable链接来增强碎片化和识别.
- 在活细胞中应用交叉链接剂进行全蛋白质组分析.
主要成果:
- 成功合成和应用水溶性,细胞透的光交叉连接剂.
- 在大肠杆菌中证明了全蛋白质组的光交叉链接.
- 用残留物级分辨率阐明大肠杆菌相互作用网络的一部分.
结论:
- 已开发的光交叉链接剂在细胞蛋白全蛋白相互作用研究中是有效的.
- 这些方法使得原生环境中的蛋白相互作用网络能够在残留水平上进行分辨.
- 这种方法有望探索细胞分子社会学.
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