相关实验视频

Updated: Sep 12, 2025

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Published on: March 23, 2020

计算驱动的基化蛋白质的自上而下的质谱

Elizaveta I Shestoperova¹, Daniil G Ivanov¹, Joyce Zhong¹

¹Department of Chemistry, University of Massachusetts Amherst, Amherst, Massachusetts, USA.

bioRxiv : the preprint server for biology

|August 6, 2025

概括

这项研究介绍了UbqTop,这是一个用于质谱的计算工具,可以同时绘制蛋白质上无处不在位和链结构的地图. 这有助于我们更好地理解具有高结构分辨率的无处不在信号传递.

科学领域:

蛋白质组学是指蛋白质组学.
生物化学生物化学
分子生物学分子生物学

背景情况:

乌比基因化是一种关键的翻译后修饰,调节细胞功能.
聚比奎丁链表现出不同的拓 (链接,长度,分支),这些都决定了信号的结果.
现有的质谱法方法难以同时解决完整蛋白质上的无处不在位点和Ub链架构.

研究的目的:

开发一种综合策略,同时确定乌比奎部位和多比奎链拓.
建立一个新的框架,用于高分辨率的蛋白形状水平分析的无处不在的信号传递.

主要方法:

开发UbqTop,一个使用贝叶斯式得分的计算平台,用于从协同MS (MS2) 数据中预测Ub链拓.
与选择性Asp-N蛋白解集成,在基质消化过程中保持完整的Ub链.
应用自上而下的质谱法 (TD-MS) 进行 Ub 链架构的现场解析映射.

主要成果:

在完好无损的蛋白质基板上,证明了在同一时间确定泛素位和链拓.
成功地解决了复杂的Ub链架构,包括异构和分支链.
验证了在自由的Ub链和多重ubiquitinated蛋白质上的方法.

结论:

开发的综合战略为分析无处不在的信号提供了前所未有的结构分辨率.
这种方法提供了一个强大的新框架,用于详细的蛋白质形式水平的特征的无处不在.
能够更深入地了解各种Ub链拓的功能后果.

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Ubiquitin Chain Analysis by Parallel Reaction Monitoring

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Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

Profiling Ubiquitin and Ubiquitin-like Dependent Post-translational Modifications and Identification of Significant Alterations

Published on: November 7, 2019

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