利用点击化学分析线粒体翻译。

4区 生物学 Q3 Biochemistry, Genetics and Molecular Biology Methods in enzymology Pub Date : 2024-01-01 Epub Date: 2024-08-23 DOI:10.1016/bs.mie.2024.07.044
Roya Yousefi, Sven Dennerlein
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引用次数: 0

摘要

线粒体含有自己的基因表达机制,可合成氧化磷酸化系统的核心亚基。监测细胞空间分区内的线粒体翻译非常困难。在这里,我们描述了一种利用点击化学方法对细胞特定部分内线粒体翻译进行可视化的方法。这种方法可用于神经元等不同类型的细胞,并能利用显微镜技术以空间分辨率检测新合成的线粒体蛋白。此外,利用点击化学方法,还可通过标准 SDS-PAGE 监测线粒体翻译。通过避免使用放射性成分,所述方法为分析细胞内新合成的线粒体编码蛋白提供了新途径。
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Analysis of mitochondrial translation using click chemistry.

Mitochondria contain their own gene expression machinery, which synthesizes core subunits of the oxidative phosphorylation system. Monitoring mitochondrial translation within spatial compartments of cells is difficult. Here we describe a method to visualize mitochondrial translation within defined parts of cells, using a click chemistry approach. This method can be applied to different cell types such as neurons and allows detection of newly synthesized mitochondrial proteins in spatial resolution using microscopy techniques. Furthermore, using click chemistry, mitochondrial translation can also be monitored by standard SDS-PAGE. The described method avenues the analysis of newly synthesized mitochondrial encoded proteins in the cellular context, by avoiding the usage of radioactive components.

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来源期刊
Methods in enzymology
Methods in enzymology 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
308
审稿时长
3-6 weeks
期刊介绍: The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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