Multi-state Mechanism of Lysozyme Denaturation through the Synchronous Analysis of Raman Spectra
Multi-state Mechanism of Lysozyme Denaturation through the Synchronous Analysis of Raman Spectra
复制标题
通过拉曼光谱同步分析溶菌酶变性的多态机制
DOI：
10.1021/acs.jpcb.6b07900
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发表时间：
2016
影响因子：
3.3
通讯作者：
Yi Luo
中科院分区：
文献类型：
--
作者：
Lei Xing;Ke Lin;Xiaoguo Zhou;Shilin Liu;Yi Luo
鸡蛋溶菌酶的变性机理至今仍有争议。本研究利用拉曼光谱研究了溶菌酶的热变性和化学变性机理。在变性过程中同步记录和分析所有的拉曼谱带。发现侧基的拉曼谱带先于骨架基团的谱带发生变化。这直接揭示了溶菌酶热变性的三态机理。盐酸胍对溶菌酶的化学变性也观察到侧基的优先变化。此外，还发现溶菌酶表面基团的拉曼谱带变化先于其他基团。这表明在每一步中，化学变性剂先与蛋白质表面发生作用，再与蛋白质核心发生作用，溶菌酶的化学变性符合多态性和由外向内的机理。同步拉曼光谱研究不仅揭示了溶菌酶变性的多态机制，而且表明这种同步拉曼光谱分析是研究其他蛋白质变性机制的有力手段。
The denaturation mechanism of hen egg lysozyme is still controversial. In this study, Raman spectroscopy was employed to study the thermal and chemical denaturation mechanisms of lysozyme. All of the Raman bands were synchronously recorded and analyzed during the denaturation process. It was found that the Raman bands of the side groups changed before the bands of skeleton groups. This directly reveals the three-state mechanism of thermal denaturation of lysozyme. The preferential change of the side groups was also observed in the chemical denaturation of lysozyme by guanidine hydrochloride. Moreover, it was found that the Raman bands of the groups on the surface of lysozyme changed before those of the other groups. This indicates that the chemical denaturants interact with the protein surface before the protein core in each step and the chemical denaturation of lysozyme conforms to the multistate and outside-in mechanisms. The synchronous Raman study not only reveals the multistate mechanism of lysozyme denaturation but also demonstrates that this synchronous Raman analysis is a powerful method to study the denaturation mechanisms of other proteins.