用共定位单分子显微镜观察细胞机器。
Visualizing cellular machines with colocalization single molecule microscopy.
机构信息
Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Dr., Madison, USA.
出版信息
Chem Soc Rev. 2014 Feb 21;43(4):1189-200. doi: 10.1039/c3cs60208g. Epub 2013 Aug 22.
Abstract
Many of the cell's macromolecular machines contain multiple components that transiently associate with one another. This compositional and dynamic complexity presents a challenge for understanding how these machines are constructed and function. Colocalization single molecule spectroscopy enables simultaneous observation of individual components of these machines in real-time and grants a unique window into processes that are typically obscured in ensemble assays. Colocalization experiments can yield valuable information about assembly pathways, compositional heterogeneity, and kinetics that together contribute to the development of richly detailed reaction mechanisms. This review focuses on recent advances in colocalization single molecule spectroscopy and how this technique has been applied to enhance our understanding of transcription, RNA splicing, and translation.
摘要
许多细胞的大分子机器都包含多个瞬时相互作用的组件。这种组成和动态的复杂性给理解这些机器的构建和功能带来了挑战。共定位单分子光谱学能够实时观察这些机器的各个组件，并为通常在整体分析中被掩盖的过程提供独特的视角。共定位实验可以提供关于组装途径、组成异质性和动力学的有价值信息，这些信息共同有助于丰富详细的反应机制的发展。本文综述了共定位单分子光谱学的最新进展，以及该技术如何被应用于提高我们对转录、RNA 剪接和翻译的理解。
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本文引用的文献
1
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Nat Struct Mol Biol. 2013 Jun;20(6):718-27. doi: 10.1038/nsmb.2567. Epub 2013 Apr 28.
2
Single-molecule colocalization FRET evidence that spliceosome activation precedes stable approach of 5' splice site and branch site.单分子 colocalization FRET 证据表明剪接体的激活先于 5' 剪接位点和分支位点的稳定接近。
Proc Natl Acad Sci U S A. 2013 Apr 23;110(17):6783-8. doi: 10.1073/pnas.1219305110. Epub 2013 Apr 8.
3
High-resolution cryo-electron microscopy structure of the Trypanosoma brucei ribosome.布鲁氏锥虫核糖体的高分辨率冷冻电子显微镜结构。
Nature. 2013 Feb 21;494(7437):385-9. doi: 10.1038/nature11872. Epub 2013 Feb 10.
4
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Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1285-90. doi: 10.1073/pnas.1211164110. Epub 2013 Jan 4.
5
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