Probing structure and dynamics of protein assemblies by magic angle spinning NMR spectroscopy.
Probing structure and dynamics of protein assemblies by magic angle spinning NMR spectroscopy.
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DOI：
10.1021/ar300309s
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发表时间：
2013-09-17
影响因子：
18.3
通讯作者：
Polenova, Tatyana
中科院分区：
文献类型：
--
作者：
Yan, Si;Suiter, Christopher L.;Hou, Guangjin;Zhang, Huilan;Polenova, Tatyana
在活的生物体中，生物分子经常组织成多组分的复合体。这些组件由各种蛋白质组成，执行从细胞分裂、运输和能量转导到催化、信号和病毒感染性的各种基本功能。为了了解这些组件在健康和疾病状态下的生物学功能，研究人员需要研究它们的三维结构和分子动力学。到目前为止，蛋白质的大尺寸，缺乏固有的长程有序性，以及不溶性，使得使用传统的结构生物学方法，如X射线衍射和溶液核磁共振光谱，对许多蛋白质组件的原子分辨研究具有挑战性或不切实际。在过去的十年里，我们的工作集中在魔角旋转固体核磁共振(MAS核磁共振)方法的开发和应用上，以表征原子水平上的大分子蛋白质组装。在这篇文章中，我们讨论了MAS核磁共振光谱学领域的快速进展，引用了我们实验室和其他实验室在方法学发展方面的工作，这些工作促进了对具有生物重要性的蛋白质组件的深入分析。我们强调提高灵敏度和分辨率的技术，例如快速MAS(40 kHz及以上的旋转频率)和用于数据采集和处理的非均匀采样协议。我们还讨论了在快速MAS条件下获得距离约束和重新耦合各向异性张量相互作用的实验。在处理蛋白质组件时，我们给出了样品制备方法的概述。在概述了当代MAS核磁共振方法之后，我们给出了我们实验室中正在研究的两类生物系统的结构和动力学的案例研究。我们首先将注意力转向细胞骨架微管马达蛋白，包括哺乳动物的动力蛋白和动力蛋白轻链8。然后我们将讨论HIV-1逆转录病毒的蛋白质组装。
In living organisms, biological molecules often organize into multi-component complexes. Such assemblies consist of various proteins and carry out essential functions, ranging from cell division, transport, and energy transduction to catalysis, signaling, and viral infectivity. To understand the biological functions of these assemblies, in both healthy and disease states, researchers need to study their three-dimensional architecture and molecular dynamics. To date, the large size, the lack of inherent long-range order, and insolubility have made atomic-resolution studies of many protein assemblies challenging or impractical using traditional structural biology methods such as X-ray diffraction and solution NMR spectroscopy. In the past ten years, we have focused our work on the development and application of magic angle spinning solid-state NMR (MAS NMR) methods to characterize large protein assemblies at atomic-level resolution. In this Account, we discuss the rapid progress in the field of MAS NMR spectroscopy, citing work from our laboratory and others on methodological developments that have facilitated the in-depth analysis of biologically important protein assemblies. We emphasize techniques that yield enhanced sensitivity and resolution, such as fast MAS (spinning frequencies of 40 kHz and above) and non-uniform sampling protocols for data acquisition and processing. We also discuss the experiments for gaining distance restraints and for recoupling anisotropic tensorial interactions under fast MAS conditions. We give an overview of sample preparation approaches when working with protein assemblies. Following the overview of contemporary MAS NMR methods, we present case studies into the structure and dynamics of two classes of biological systems under investigation in our laboratory. We will first turn our attention to cytoskeletal microtubule motor proteins including mammalian dynactin and dynein light chain 8. We will then discuss protein assemblies from the HIV-1 retrovirus.
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影响因子：
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通讯作者：
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