发光学报, 2015, 36 (12): 1458, 网络出版: 2016-02-02
硝基羟乙唑与溶菌酶反应机制的荧光光谱研究
Investigation of The Interaction Mechanism Between Lysozyme and Trichazol Using Fluorescence Spectroscopic Method
荧光光谱法 同步光谱法 溶菌酶 硝基羟乙唑 反应机理 fluorescence spectroscopy synchronous fluorescence spectroscopy lysozyme trichazol interaction mechanism
摘要
分别在298,310,318 K温度下,利用荧光光谱法研究了pH=7.40生理条件下硝基羟乙唑(TRI)与溶菌酶(LYSO)的相互作用机理。结果表明,TRI与LYSO间通过静态猝灭方式相互作用。测定了LYSO与TRI反应的结合常数、结合位点数。利用反应过程的热力学参数,确定了LYSO-TRI体系的作用力类型;由Hill系数得出了LYSO或TRI的协同性;根据非辐射能量转移理论,确定了TRI到LYSO的结合距离,同时采用同步光谱法考察了TRI对LYSO构象的影响。
Abstract
The interaction mechanism of trichazol (TRI) to lysozyme(LYSO) at different temperatures (298, 310, 318 K) was investigated using fluorescence spectroscopy under simulative physiological conditions. The results clearly demonstrated that TRI caused strong quenching of the fluorescence of LYSO by a static quenching mechanism. The binding constants were order of magnitude of 104 and the number of binding site in the interaction was closed to 1. The thermodynamic parameters of LYSO-TRI system were determined as follows: negative ΔH and positive ΔS, which indicated that electrostatic interaction played a major role in the binding reaction. The effect of cooperative binding of TRI was quantified by Hills coefficient (nH) and the values of nH were slightly more than 1 in the systems, which indicated weakly positive cooperativeness in the interaction of TRI with LYSO. The binding distance (r) between TRI and LYSO was obtained based on the Frster nonradioactive resonance energy transfer and r was less than 7 nm. Furthermore, the effect of TRI on the conformation of LYSO was analyzed by using the synchronous fluorescence spectroscopy.
刘保生, 韩荣, 李志云, 李彤彤, 李改霞. 硝基羟乙唑与溶菌酶反应机制的荧光光谱研究[J]. 发光学报, 2015, 36(12): 1458. LIU Bao-sheng, HAN Rong, LI Zhi-yun, LI Tong-tong, LI Gai-xia. Investigation of The Interaction Mechanism Between Lysozyme and Trichazol Using Fluorescence Spectroscopic Method[J]. Chinese Journal of Luminescence, 2015, 36(12): 1458.