荧光光谱法研究盐酸阿霉素与溶菌酶的相互作用
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河北大学生命科学学院,河北大学生命科学学院

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Q55;TN249

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]保定市科学技术研究与发展指导计划(15ZN001);河北省高等学校科学技术研究项目(QN2015174);河北大学中西部高校提升综合实力工程项目资助;河北省生物学强势特色学科建设项目


Studies on the Interaction of Lysozyme with Doxorubicin hydrochloride by Fluorescence Spectroscopy
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    摘要:

    盐酸阿霉素(Doxorubicin hydrochloride, DOX)是临床上广泛应用的一种抗癌药物,溶菌酶(Lysozyme, LYS)是生物体内普遍存在的一种具有杀菌、抗病毒、抗肿瘤作用的蛋白质,研究DOX与LYS之间的相互作用十分必要。本研究利用荧光光谱法研究了不同温度下(298 K、308 K 和318 K)DOX与LYS的相互作用,阐述了DOX对LYS荧光猝灭的机理,并计算了二者之间的结合位点和结合常数。结果表明,DOX能使LYS的内源荧光发生猝灭,荧光猝灭机理属于二者形成复合物所引起的静态猝灭。Stern-Volmer方程分析显示,DOX与LYS具有1个结合位点、不同温度下DOX与LYS的结合常数(KA):4.79×106 (298 K)、2.82×106 (308 K)和2.14×106(318 K)。通过计算热力学参数,可知DOX与LYS的相互作用主要是由分子之间的静电作用引发的,并且二者之间相互作用是伴随吉布斯自由能降低的自发反应。三维荧光光谱实验显示,DOX的加入引起LYS构象的变化,表现为蛋白质内部色氨酸残基所处微环境的疏水性降低。

    Abstract:

    Doxorubicin hydrochloride (DOX) is a kind of medicine with widely application of anti-cancer. Lysozyme (LYS), a protein existing in organism, has effect of anti-bacteria, anti-virus, and antineoplastic. Thus, Studies on the Interaction of LYS with DOX is necessary. The interactions between DOX and LYS at different temperature were studied by fluorescence spectroscopy. The quenching mechanism between DOX and LYS was demonstrated, and the binding parameters were determined. The results showed that the intrinsic fluorescence of LYS could be quenched by DOX. The quenching mechanism between is due to static manner by forming DOX-LYS complex. The binding parameters were determined according to Stern–Volmer equation, and the thermodynamic parameters were calculated. The results showed, there is one binding site between DOX and LYS. The binding constants (KA) between DOX and LYS at different temperature were 4.79×106 (298 K), 2.82×106 (308 K), and 2.14×106 (318 K), respectively. Thermodynamic analysis indicated that the interaction process was spontaneous, and electrostatic force might be primarily responsible for the interaction. In addition, the effect of DOX on the conformation of LYS was analyzed by three-dimensional fluorescence spectra. The result indicates that the polarity microenvironment around Trp residues decreased.

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倪志华,张玉明. 荧光光谱法研究盐酸阿霉素与溶菌酶的相互作用[J]. 科学技术与工程, 2016, 16(5): .
NI Zhi-hua and. Studies on the Interaction of Lysozyme with Doxorubicin hydrochloride by Fluorescence Spectroscopy[J]. Science Technology and Engineering,2016,16(5).

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  • 收稿日期:2015-10-10
  • 最后修改日期:2016-01-27
  • 录用日期:2015-11-26
  • 在线发布日期: 2016-02-22
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