3D protein structures by solid-state NMR spectroscopy:: Ready for high resolution
3D protein structures by solid-state NMR spectroscopy:: Ready for high resolution
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DOI：
10.1002/anie.200801352
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发表时间：
2008-01-01
影响因子：
16.6
通讯作者：
Bockmann, Anja
中科院分区：
文献类型：
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作者：
Bockmann, Anja
高分辨率蛋白质三维结构的解决方案是结构生物学的一项重大成就。蛋白质结构是详细了解蛋白质功能的基础;酶如何进行化学反应;分子如何通过膜运输;以及如何施加机械力。它们允许识别三维蛋白质结构中的关键基序，并因此允许开发策略，通过设计抑制剂来破坏蛋白质功能，并产生突变体来回答详细的问题。3D结构形成了其他结构研究的先决条件，例如研究蛋白质-蛋白质相互作用或与核酸，脂质或溶剂的相互作用，并且还允许蛋白质动力学在空间中映射。没有3D结构，这种蛋白质动力学的理解不能被翻译成空间分辨的information.Magic-angle-spinning（MAS）的固体核磁共振实验的结构测定均匀13 C，15 N同位素富集蛋白质最近经历了重大进展，该方法是连接X射线晶体学和液态核磁共振光谱学作为一种工具，结构生物学。固体核磁共振波谱主要用于研究不溶性蛋白质的结构，如纤维和膜蛋白。这些蛋白质在蛋白质数据库中的代表性很差，[1]因为它们的结构很难通过其他方法获得。决定性的一步是在20世纪90年代末通过固态NMR光谱对完全13 C，15 N标记的蛋白质进行研究;与使用少量选择性标记的研究相比，这些方法可以从整个蛋白质序列中获得信息。证明使用固态NMR实验进行顺序共振分配是可能的是一个突破，并被证明可以立即用于基于这些化学位移的大量研究，包括蛋白质相互作用，动力学和折叠。
The solution of high-resolution 3D protein structures is a major achievement of structural biology. Protein structures are the basis for a detailed understanding of how a protein functions; how enzymes do chemistry; how molecules are transported through membranes; and how mechanical forces can be exerted. They allow the identification of the key motifs in three-dimensional protein structures and as such allow the development of strategies to undermine protein function through the design of inhibitors and also to produce mutants to answer detailed questions. 3D structures form the prerequisite for other structural studies—such as the investigation of protein–protein interactions or interactions with nucleic acids, lipids, or solvent—and also allow protein dynamics to be mapped in space. Without 3D structures, this understanding of protein dynamics cannot be translated into spatially resolved information.Magic-angle-spinning (MAS) solid-state NMR experiments for structure determination of uniformly 13C, 15N isotopically enriched proteins have undergone major advances recently, and the method is on the way to joining X-ray crystallography and liquid-state NMR spectroscopy as a tool for structural biology. Solid-state NMR spectroscopy mainly aims at structural studies of insoluble proteins, such as fibrils and membrane proteins. These proteins are poorly represented in the Protein Data Bank,[1] as their structures are difficult to obtain by other methods. A decisive step was the development of studies on fully 13C, 15N-labeled proteins by solid-state NMR spectroscopy at the end of the 1990s; in contrast to studies using few selective labels, these approaches allow information to be obtained from the entire protein sequence. The proof that sequential resonance assignments using solid-state NMR experiments are possible was a breakthrough and proved to be immediately useful for a plethora of studies based on these chemical shifts including protein interactions, dynamics, and folding.