多光谱法研究溶菌酶不对称变复性机理  

Studies on the asymmetric mechanism of Lysozyme's unfolding and refolding using multi-spectroscopic method

作　　者：高小艳[1,2] 郭明[2] 

机构地区：[1]塔里木大学生命科学学院,新疆阿拉尔843300 [2]浙江农林大学理学院,浙江临安311300

出　　处：《分子科学学报》2014年第6期470-480,共11页Journal of Molecular Science

基　　金：浙江省科技计划资助项目(2012R412052)

摘　　要：利用紫外-可见光谱法、荧光相图法、荧光偏振法以及共振瑞利散射法多光谱技术对溶菌酶(Lysozyme)的变复性特征进行表征,分析溶菌酶变复性机理,构建其变性动力学模型.结果表明,变性过程溶菌酶分子体积变大,结构松弛,变性速度快,不存在中间态,符合"二态模型";复性过程变性溶菌酶分子体积变小,结构紧缩,复性过程慢,且存在中间体,符合"多态模型",说明了溶菌酶的复性过程比其变性过程复杂.实验定量研究了不同变复性条件下,溶菌酶不对称变复性机理,为溶菌酶分子结构分析的研究提供了参考.UV-visible spectroscopy, fluorescence phase diagram, fluorescence polarization and Reso- nance Rayleigh Scattering （RRS） methods were used for characterizing the complex features of unfol- ding lysozyme and refolding lysozyme. What＇s more, the mechanisms were analyzed, and its unfol- ding kinetic model was built. The results showed that the volumes of lysozyme molecule enlarged, its structural got slack, the speed of its molecule motion became faster, and there was no intermediate state in the process of denaturation lysozyme, which conformed to ＂two-state model＂. It was opposite to process of denaturation lysozyme in the process of refolding, both the structure and the volumes of lysozyme molecule were decreased, In addition, the refolding process was slow, and there was an in- termediate state. This process was line with ＂multi-state model＂. It described that the process of re- folding of lysozyme is more complex than its denaturation process. The asymmetry mechanism of lyso- zyme under different conditions, including unfolding system and refolding system, was quantitatively studied in this experiment, it provided a reference for the study of the molecular structure of lyso- zyme.

关 键 词：溶菌酶 多光谱法 机理 变复性 动力学模型 

分 类 号：O654[理学—分析化学]