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完整噬菌体的魔角旋转核磁共振:对衣壳、DNA及其界面的深入了解

Magic-angle spinning NMR of intact bacteriophages: insights into the capsid, DNA and their interface.

作者信息

Abramov Gili, Morag Omry, Goldbourt Amir

机构信息

School of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel Aviv University, Ramat Aviv 69978, Tel Aviv, Israel.

School of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel Aviv University, Ramat Aviv 69978, Tel Aviv, Israel.

出版信息

J Magn Reson. 2015 Apr;253:80-90. doi: 10.1016/j.jmr.2015.01.011.

Abstract

Bacteriophages are viruses that infect bacteria. They are complex macromolecular assemblies, which are composed of multiple protein subunits that protect genomic material and deliver it to specific hosts. Various biophysical techniques have been used to characterize their structure in order to unravel phage morphogenesis. Yet, most bacteriophages are non-crystalline and have very high molecular weights, in the order of tens of MegaDaltons. Therefore, complete atomic-resolution characterization on such systems that encompass both capsid and DNA is scarce. In this perspective article we demonstrate how magic-angle spinning solid-state NMR has and is used to characterize in detail bacteriophage viruses, including filamentous and icosahedral phage. We discuss the process of sample preparation, spectral assignment of both capsid and DNA and the use of chemical shifts and dipolar couplings to probe the capsid-DNA interface, describe capsid structure and dynamics and extract structural differences between viruses.

摘要

噬菌体是感染细菌的病毒。它们是复杂的大分子组装体,由多个蛋白质亚基组成,这些亚基保护基因组物质并将其传递给特定宿主。为了阐明噬菌体的形态发生,人们使用了各种生物物理技术来表征其结构。然而,大多数噬菌体是非晶体的,分子量非常高,达到数十兆道尔顿。因此,对包含衣壳和DNA的此类系统进行完整的原子分辨率表征非常罕见。在这篇观点文章中,我们展示了魔角旋转固态核磁共振如何以及如何被用于详细表征噬菌体病毒,包括丝状噬菌体和二十面体噬菌体。我们讨论了样品制备过程、衣壳和DNA的光谱归属以及使用化学位移和偶极耦合来探测衣壳-DNA界面,描述衣壳结构和动力学,并提取病毒之间的结构差异。

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