核磁共振波谱法在蛋白质三维结构解析中的应用

蛋白质特定的三维结构与其生物功能密切相关，因此，研究蛋白质的三维结构有助于揭示其生物功能机制。将核磁共振（NMR）波谱法应用于研究溶液状态下蛋白质的三维结构，能够更加准确地揭示蛋白质结构与生物功能之间的关系。本文综述了NMR解析蛋白质三维结构的理论和技术方法，以及NMR结合其他生物物理手段，并辅以分子建模计算法研究蛋白质三维结构的研究进展和最新方法，为精准解析蛋白质的三维结构提供思路及策略。

Application of Nuclear Magnetic Resonance Spectroscopy for Studying Protein Three-dimensional Structure

The unique three-dimensional structure of protein is closely related to its biological function. Therefore, investigating the three-dimensional structure of protein is helpful to reveal its biological function mechanism. The study of protein three-dimensional structure in the solution state using nuclear magnetic resonance (NMR) spectroscopy can accurately reveal the relationship between protein structure and biological function. The aim of this article is to provide an effective strategy for accurate analysis of protein three-dimensional structure using NMR and combination with other biophysical means such as molecular modeling and computation methods through reviewing the research progress and latest technology in these fields. Firstly, we summarize the theory of NMR for studying protein three-dimensional structure. Secondly, we deeply review the theory and technology of NMR analysis of protein three-dimensional structure, including isotope labeling of proteins (labeling methods, expression systems, and purification techniques), NMR data acquisition and analysis software, analysis of amino acid sequence, secondary structural unit and three-dimensional structure of protein using NMR and combination with other biophysical means (F?rster/fluorescence resonance energy transfer (FRET), chemical cross-linking coupled with mass spectrometry (CXMS), small angel X-ray scattering (SAXS), and cryo-electron microscopy (Cryo-EM)), and analysis of excited state structure of protein molecules using Carr-Purcell-Meiboom-Gill relaxation dispersion (CPMG RD) and chemical exchange saturation transfer (CEST) techniques. Thirdly, we overview recent researches about application of NMR for analysis of three-dimensional structure of high molecular mass single chain protein and supramolecular protein complex. Fourthly, we elaborate the latest progress in the field of NMR combined with molecular modeling and computation methods. Lastly, we summarize challenges and prospects of application of NMR for studying protein three-dimensional structure in the future.