Mechanistic insights into antibiotic action on the ribosome through single-molecule fluorescence imaging.

Mechanistic insights into antibiotic action on the ribosome through single-molecule fluorescence imaging.
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通过单分子荧光成像对核糖体对抗生素作用的机理见解。

DOI:
10.1111/j.1749-6632.2012.06839.x
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发表时间:
2011-12
影响因子:
5.2
通讯作者:
Blanchard SC
Blanchard SC
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Wang L;Wasserman MR;Feldman MB;Altman RB;Blanchard SC

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单分子荧光成像为核糖体功能动力学提供了前所未有的途径,揭示了对核糖体活性至关重要的瞬态中间状态。现在已经开发出能够同时探测数百个分子的成像平台,其时间和空间分辨率接近亚毫秒时间和亚纳米尺度。这些进步使得在翻译过程中的单个步骤以及过程反应的稳定和预稳定状态测量成为可能。使用这些方法产生的数据产生了新的,定量的结构和动力学的见解核糖体的活性。他们还揭示了抗生素靶向翻译装置的机制,揭示了通过其他方法难以获得的结构-功能关系的特征。这篇综述概述了使用这种成像平台可以获得的信息类型,并提供了使用该技术评估小分子抗生素如何改变大分子功能的蓝图。
Single-molecule fluorescence imaging has provided unprecedented access to the dynamics of ribosome function, revealing transient intermediate states that are critical to ribosome activity. Imaging platforms have now been developed that are capable of probing many hundreds of molecules simultaneously at temporal and spatial resolutions approaching the sub-millisecond time and the sub-nanometer scales. These advances enable both steady- and pre-steady state measurements of individual steps in the translation process as well as processive reactions. The data generated using these methods have yielded new, quantitative structural and kinetic insights into ribosomal activity. They have also shed light on the mechanisms of antibiotics targeting the translation apparatus, revealing features of the structure-function relationship that would be difficult to obtain by other means. This review provides an overview of the types of information that can be obtained using such imaging platforms and a blueprint for using the technique to assess how small-molecule antibiotics alter macromolecular functions.
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