相关实验视频
Updated: May 5, 2026

09:45
In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
8.8K
对Mdm2的SUMO-1修改阻止了其自我无处置,并增加了Mdm2对p53无处置的能力
T Buschmann1, S Y Fuchs, C G Lee
1Derald H Ruttenberg Cancer Center, Mount Sinai School of Medicine, New York, New York 10029, USA.
Cell
|July 13, 2000
概括
在Lys-446的Mdm2 E3无素酶化对p53调节至关重要. 降低Mdm2相化增强了p53稳定性和DNA损伤后的亡.
科学领域:
- 分子生物学分子生物学
- 癌症研究 癌症研究
- 蛋白质生物化学 蛋白质生物化学
背景情况:
- Mdm2 是一个关键的E3泛素酶,向p53瘤抑制剂.
- 调节Mdm2活动对于控制p53水平和功能至关重要.
- 翻译后的修改,如sumoylation,可以调节E3酶活性.
研究的目的:
- 研究SUMO-1结合 (sumoylation) 在Mdm2.2上的作用.
- 确定Mdm2相化对其对p53.3的活性所带来的功能影响.
- 探索DNA损伤对Mdm2相和p53稳定性的影响.
主要方法:
- 局部定向的突变发生产生Mdm2 ((K446R) 突变.
- 在Mdm2.2.的体外合试验.
- 分析Mdm2的自我无处置和p53的无处置.
- 西部涂抹测试以评估Mdm2和p53.5的蛋白质水平.
- 辐射实验用于诱导DNA损伤.
主要成果:
- 在RING指域内,Mdm2在Lys-446中被sumoylated.
- Mdm2 ((K446R) 突变显示稳定性增加,但增强了p53降解.
- 在Mdm2的体外结合抑制了自我ubiquitination,并增强了p53酶的活性.
- 辐射降低了Mdm2的sumoylation,与p53水平相反相关.
结论:
- Mdm2 E3结合酶的活性依赖于sumoylation来维持其内在的功能.
- 在DNA损伤时减少Mdm2的sumoylation有助于p53的稳定.
- Mdm2的sumoylation代表了p53通路激活的新型调节机制.
相关概念视频
Negative Regulator Molecules
32.1K
Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.
32.1K
Covalently Linked Protein Regulators
8.2K
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....
8.2K
Master Transcription Regulators
6.1K
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
6.1K
Abnormal Proliferation
4.0K
Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the...
4.0K
mTOR Signaling and Cancer Progression
3.6K
The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
The mTOR pathway or the...
3.6K
Interactions Between Signaling Pathways
4.7K
Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
4.7K

