相关实验视频
Updated: Sep 17, 2025

08:33
Ubiquitin Chain Analysis by Parallel Reaction Monitoring
Published on: June 17, 2020
3.7K
在细胞中使用协同存在的ubiquitin结合实体来解链特定的ubiquitination
Muhammad Shahzad Ali1, Christopher Rainville1, Janelle Pedroza2
1Progenra Inc, 271A Great Valley Parkway, Malvern, PA, 19335, USA.
Scientific reports
|July 2, 2025
概括
这项研究引入了链选择性并列乌比基因结合实体 (TUBE),用于对蛋白质乌比基因化结合类型的高通量评估. 该方法成功地区分了RIPK2的K48和K63无处不在,推动了向蛋白质降解研究.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 细胞信号传输 细胞信号传输
背景情况:
- 聚尤比基因化通过独特的乌比基因链接来调节多种细胞功能.
- 氨酸48 (K48) 连接链针对蛋白质的蛋白质分解,而氨酸63 (K63) 连接链参与信号和贩运.
- 无素-蛋白酶体系统 (UPS) 是药物发现的目标,蛋白质溶解向金马 (PROTACs) 和分子粘剂 (MGs) 劫持E3酶以进行向蛋白质降解.
研究的目的:
- 开发和验证一种高通量方法,用于评估内源蛋白的链接特异性无化.
- 以一种链接特定的方式研究RIPK2的泛化动态,这是一个关键的炎症信号调节器.
主要方法:
- 应用特定链的并列乌比基因结合实体 (TUBE),对聚比基因链有很高的亲和力.
- 使用高通量选 (HTS) 试验来分析无处不在的模式.
- 采用L18-MDP诱导K63的泛化,并使用RIPK2 PROTAC诱导RIPK2的K48泛化.
主要成果:
- 证明了链选择性TUE能够在K48和K63之间区分内源性RIPK2的无处不在2.
- 成功监测了RIPK2对特定刺激的反应中的链接特异性泛化动态.
- 验证的TUBE作为一种工具,以依赖于上下文的方式对蛋白质无化进行高通量评估.
结论:
- 链选择性管道为高通量,链接特定的无处不在分析提供了强大的平台.
- 这项技术使人们能够更深入地了解各种细胞环境中的泛素代码.
- 开发的方法有可能在药物发现和理解信号通路方面得到更广泛的应用.
相关概念视频
Regulated Protein Degradation
7.7K
It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
7.7K
Protein Complexes with Interchangeable Parts
2.6K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.6K
The Proteasome
1.2K
Eukaryotic cells can degrade proteins through several pathways. One of the most important among these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
1.2K
Covalently Linked Protein Regulators
7.4K
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein....
These groups modify specific amino acids in a protein....
7.4K
The Proteasome Structure
990
The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
The proteasome is an...
The proteasome is an...
990
Anaphase Promoting Complex
3.0K
The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
3.0K

