Yang Shouning, Zhang Qiaoling, Yang Huayan, Shi Haimei, Dong Aichun, Wang Li, Yu Shaoning
Zhejiang Provincial Key Laboratory of Advanced Mass Spectrometry and Molecular Analysis, Institute of Mass Spectrometry, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, Zhejiang 315211, China.
Kweichow Moutai Group, Renhuai, Guizhou 564501, China.
Int J Biol Macromol. 2022 May 1;206:175-187. doi: 10.1016/j.ijbiomac.2022.02.104. Epub 2022 Feb 22.
Infrared (IR) spectroscopy is a highly sensitive technique that provides complete information on chemical compositions. The IR spectra of proteins or peptides give rise to nine characteristic IR absorption bands. The amide I bands are the most prominent and sensitive vibrational bands and widely used to predict protein secondary structures. The interference of HO absorbance is the greatest challenge for IR protein secondary structure prediction. Much effort has been made to reduce/eliminate the interference of HO, simplify operation steps, and increase prediction accuracy. Progress in sampling and equipment has rendered the Fourier transform infrared (FTIR) technique suitable for determining the protein secondary structure in broader concentration ranges, greatly simplifying the operating steps. This review highlights the recent progress in sample preparation, data analysis, and equipment development of FTIR in A/T mode, with a focus on recent applications of FTIR spectroscopy in the prediction of protein secondary structure. This review also provides a brief introduction of the progress in ATR-FTIR for predicting protein secondary structure and discusses some combined IR methods, such as AFM-based IR spectroscopy, that are used to analyze protein structural dynamics and protein aggregation.
红外（IR）光谱法是一种高度灵敏的技术，可提供有关化学成分的完整信息。蛋白质或肽的红外光谱会产生九个特征性红外吸收带。酰胺I带是最突出且最灵敏的振动带，广泛用于预测蛋白质二级结构。HO吸光度的干扰是红外蛋白质二级结构预测面临的最大挑战。人们已付出诸多努力来减少/消除HO的干扰、简化操作步骤并提高预测准确性。采样和设备方面的进展使傅里叶变换红外（FTIR）技术适用于在更宽的浓度范围内测定蛋白质二级结构，极大地简化了操作步骤。本综述重点介绍了衰减全反射（ATR）模式下FTIR在样品制备、数据分析和设备开发方面的最新进展，尤其关注FTIR光谱在蛋白质二级结构预测中的最新应用。本综述还简要介绍了衰减全反射傅里叶变换红外光谱（ATR-FTIR）在预测蛋白质二级结构方面的进展，并讨论了一些用于分析蛋白质结构动力学和蛋白质聚集的联合红外方法，如基于原子力显微镜（AFM）的红外光谱法。
