A Chemical Biology Primer for NMR Spectroscopists.
A Chemical Biology Primer for NMR Spectroscopists.
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DOI：
10.1016/j.jmro.2022.100044
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发表时间：
2022-02
影响因子：
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通讯作者：
Evan T. Clark;Elanor E. Sievers;G. Debelouchina
中科院分区：
文献类型：
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作者：
Evan T. Clark;Elanor E. Sievers;G. Debelouchina
在结构生物学技术中，核磁共振波谱提供了独特的能力，使动态和异质生物系统在生理和自然条件下的原子分辨率研究成为可能。然而，复杂的生物系统往往以其低灵敏度、拥挤的光谱或反映其复杂相互作用模式和动力学的大线宽挑战核磁共振波谱学家。虽然这些挑战可以通过新的光谱方法的发展来克服，但化学生物学也可以在样品制备阶段提供优雅而有效的解决方案。在本教程中，我们旨在介绍几种化学生物学工具，这些工具可以制备选择性和分段标记的蛋白质样品，以及介绍位点特异性光谱探针和翻译后修饰。本文所涉及的四种工具，即半胱氨酸化学、内部链、天然化学连接和非天然氨基酸结合，近年来得到了发展和优化，比以往任何时候都更有效，适用于更广泛的蛋白质。我们简要介绍了每种工具，描述了其在核磁共振实验背景下的优缺点，并为样品制备和分析提供了实用的建议。我们希望本教程将向该领域的初级研究人员介绍化学生物学可以为核磁共振波谱学家提供的可能性，并将在寻求了解蛋白质在健康和疾病中的功能方面激发新的令人兴奋的应用。
Among structural biology techniques, NMR spectroscopy offers unique capabilities that enable the atomic resolution studies of dynamic and heterogeneous biological systems under physiological and native conditions. Complex biological systems, however, often challenge NMR spectroscopists with their low sensitivity, crowded spectra or large linewidths that reflect their intricate interaction patterns and dynamics. While some of these challenges can be overcome with the development of new spectroscopic approaches, chemical biology can also offer elegant and efficient solutions at the sample preparation stage. In this tutorial, we aim to present several chemical biology tools that enable the preparation of selectively and segmentally labeled protein samples, as well as the introduction of site-specific spectroscopic probes and post-translational modifications. The four tools covered here, namely cysteine chemistry, inteins, native chemical ligation, and unnatural amino acid incorporation, have been developed and optimized in recent years to be more efficient and applicable to a wider range of proteins than ever before. We briefly introduce each tool, describe its advantages and disadvantages in the context of NMR experiments, and offer practical advice for sample preparation and analysis. We hope that this tutorial will introduce beginning researchers in the field to the possibilities chemical biology can offer to NMR spectroscopists, and that it will inspire new and exciting applications in the quest to understand protein function in health and disease.