相关实验视频
Updated: May 22, 2025
10:06
Cell Type-specific Gene Expression Profiling in the Mouse Liver
Published on: September 17, 2019
7.6K
使用TurboID-Cp和OGAM对Drosophila中的O-GlcNAcylated蛋白进行组织特异性分析
Qin Lei1, Haibin Yu1, Fang Chen1
1Hunan Key Laboratory of Molecular Precision Medicine, Department of Oncology, Xiangya Hospital, Central South University, Changsha, China.
Bio-protocol
|March 14, 2025
概括
研究人员开发了一种新的工具,它结合了修饰的酶和近距离标记,以识别特定组织中的O-GlcNAcylated蛋白质. 这种方法有助于理解O-GlcNAcylation的理解.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 后翻译修改 后翻译修改
背景情况:
- 蛋白质O-GlcNAcylation是一种动态的翻译后修饰,对于转录和蛋白质平衡等细胞过程至关重要.
- O-GlcNAc 稳态失调与包括心血管,癌症和神经退行性疾病在内的疾病有关.
- 对O-GlcNAcylated蛋白质的组织特异性鉴定对于疾病发病研究至关重要,但仍然具有挑战性.
研究的目的:
- 开发一种用于O-GlcNAcylated蛋白质的组织特异性丰富和识别的新工具.
- 为了研究O-GlcNAcylation在特定组织中的作用及其与疾病发病的联系.
- 为剖析特定组织的O-GlcNAcylation功能提供全面的工作流程.
主要方法:
- 开发一种新型工具,将一种催化不活的O-GlcNAcase (CpOGA) 突变 (CpOGACD) 与TurboID近距离标记技术相结合.
- 使用TurboID-CpOGADM作为对照来区分O-GlcNAc-独立的蛋白质-蛋白质相互作用.
- 整合了Drosophila中的UAS/Gal4系统,用于针对性表达和O-GlcNAcylated蛋白质的特定组织丰富.
主要成果:
- 成功将O-GlcNAc修饰转化为生物素标签,使O-GlcNAcylated蛋白质的丰富成为可能.
- 启用质谱 (MS) 识别在特定的Drosophila组织中的O-GlcNAcylated蛋白质.
- 证明了该工具在以特定组织的方式丰富O-GlcNAc蛋白质的有效性.
结论:
- 开发的TurboID-CpOGAM工具为研究O-GlcNAcylation提供了一种创新的方法.
- 这种方法促进了O-GlcNAcylated基质的组织特异性丰富和识别.
- 为剖析组织特异性O-GlcNAcylation在健康和疾病中的功能作用提供了宝贵的资源.
相关概念视频
01:30General Transcription Factors
5.1K
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
5.1K
02:24Ribosome Profiling
3.4K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.4K
02:16Transcription Factors
75.5K
Tissue-specific transcription factors contribute to diverse cellular functions in mammals. For example, the gene for beta globin, a major component of hemoglobin, is present in all cells of the body. However, it is only expressed in red blood cells because the transcription factors that can bind to the promoter sequences of the beta globin gene are only expressed in these cells. Tissue-specific transcription factors also ensure that mutations in these factors may impair only the function of...
75.5K