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高分辨单颗粒分析在生物样品中的应用

Application of high resolution single particle analysis by cryo-electron microscopy in biological samples

  • 摘要: 冷冻电子显微学作为结构生物学主要工具之一,以解析承担生命活动中重要功能的各种蛋白质机器三维结构为手段,对其在生命活动中的功能机制进行研究。冷冻电子显微学的单颗粒分析法具有分辨率高、生物样品无需结晶、解析的结构接近生理状态等优点。近几年来,随着软、硬件的发展与革新,高分辨冷冻电镜单颗粒分析法在生物样品上的应用范围方面得到进一步扩展,囊括大至上百纳米的病毒颗粒,小至十纳米左右的蛋白质机器,同时单颗粒分析的分辨率也在稳步提高。目前,大量生物样品的结构解析能够常态地达到3~4 Å的准原子分辨率,足以阐明大部分生物问题。文章简要介绍了单颗粒分析的理论基础、发展过程、研究现状以及面临的更多挑战。

     

    Abstract: Cryo-electron microscopy (cryo-EM), a major tool in structure biology, is used to reveal the three dimensional structures of important functional protein machineries.Since crystallization is not required, sample preparation in cryo-EM is less complicated and the sample is preserved in a more native state compared with other tools in structure biology such as X-ray crystallography. Recent progress in hardware and software development of this technique has not only improved the resolution of the reconstructions but also increased the range of applications from viral particles with a diameter above 100 nm to protein complexes with a diameter around 10 nm. Now, a 3~4 Å resolution can be regularly achieved, which is adequate for answering most biological questions. In this review we describe the theoretical basis as well as the current progress of high resolution single particle analysis. The remaining challenges are also discussed.

     

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