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河北大学化学与环境科学学院 河北省分析科学技术重点实验室,河北 保定,071002
纸质出版日期:2015-12-10,
收稿日期:2015-6-21,
修回日期:2015-10-27,
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刘保生, 韩荣, 李志云等. 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): 1458-1463
LIU Bao-sheng, HAN Rong, LI Zhi-yun etc. Investigation of The Interaction Mechanism Between Lysozyme and Trichazol Using Fluorescence Spectroscopic Method[J]. Chinese Journal of Luminescence, 2015,36(12): 1458-1463
刘保生, 韩荣, 李志云等. 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): 1458-1463 DOI: 10.3788/fgxb20153612.1458.
LIU Bao-sheng, HAN Rong, LI Zhi-yun etc. Investigation of The Interaction Mechanism Between Lysozyme and Trichazol Using Fluorescence Spectroscopic Method[J]. Chinese Journal of Luminescence, 2015,36(12): 1458-1463 DOI: 10.3788/fgxb20153612.1458.
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。
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 10
4
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
H
and positive
S
which indicated that electrostatic interaction played a major role in the binding reaction. The effect of cooperative binding of TRI was quantified by Hill's coefficient (
n
H
) and the values of
n
H
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.
荧光光谱法同步光谱法溶菌酶硝基羟乙唑反应机理
fluorescence spectroscopysynchronous fluorescence spectroscopylysozymetrichazolinteraction mechanism
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