光谱学与光谱分析, 2018, 38 (3): 990, 网络出版: 2018-04-09  

光谱法和分子对接方法研究罗利环素与人血清白蛋白的相互作用

The Interaction between Rolitetracycline and Human Serum Albumin Using Multi-Spectral Methods and Molecular Modeling
作者单位
1 中央民族大学生命与环境科学学院, 北京 100081
2 中央民族大学北京市食品环境与健康工程技术研究中心, 北京 100081
摘要
在模拟人体生理条件下, 应用多光谱法和分子对接技术对罗利环素(RTC)与人血清白蛋白(HSA)的相互作用进行了研究。 实验通过消除内滤光来提高荧光数据的准确性。 此外, 荧光光谱和紫外光谱的结果表明RTC与HSA的猝灭方式是静态猝灭, 且在298和310 K温度下的结合位点数分别为1.09和0.95。 在两个不同温度下的结合常数分别为K298 K=3.13×105 L·mol-1和K310 K=0.70×105 L·mol-1。 根据热力学参数的计算结果可知, RTC与HSA的结合作用力主要是氢键和范德华力。 取代实验表明, RTC在HSA的Site Ⅰ, ⅡA子域上有一个结合位点。 根据非辐射能量转移理论得到的RTC和HSA氨基酸残基间的结合距离为2.59 nm。 三维荧光光谱表明RTC和HSA的相互作用改变了蛋白质的构象。 此外, 采用傅里叶变换红外光谱法对RTC与HSA作用前后HSA二级结构的变化进行了定量分析, 结果表明, α-螺旋结构含量降低了8.4%, β-折叠含量从30.3%增加到31.4%, β-转角也从15.6%增加到了16.1%。 分子对接进一步显示RTC通过氢键、 疏水作用力、 极性键等多种作用力与HSA的ⅡA子域上氨基酸残基相互作用。 实验结果有助于从分子水平研究RTC和HSA的相互作用。
Abstract
The interaction between rolitetracycline (RTC) and human serum albumin (HSA) has been investigated by using multi-spectral methods and molecular modeling under physiological conditions. Fluorescence spectra results revealed the presence of static quenching mechanism in the binding of RTC to HSA. The binding constants were of K298 K=3.13×105 L·mol-1 and K310 K=0.70×105 L·mol-1, respectively. The numbers of binding site were 1.09 and 0.95 at two different temperatures. The thermodynamic parameters indicated that hydrogen bond and van der Waals force were the major driving forces for interaction between RTC and HSA. The binding distance of RTC to HSA was calculated to 2.59 nm based on fluorescence resonance energy transfer. UV-Vis absorption spectra and 3-D fluorescence demonstrated that the conformation and micro-environment of HSA were changed with the addition of RTC. FT-IR spectra was used to quantitatively calculate the alternations of secondary structure of HSA with the α-helices content reduction from 51.5% to 43.1%, increasing the content of β-sheet (30.3%~31.4%) and β-turn (15.6%~16.1%). According to molecular modeling studies and site marker competitive experiments, it indicated that RTC bound in the Sudlow’s drug binding site Ⅰ of HSA and the subdomain ⅡA where Lys195, Arg218 and Arg222 residues were located. The work is helpful to understand the interaction mechanism between RTC and HSA at molecular level.

方庆, 董澄宇, 王宇, 刘颖. 光谱法和分子对接方法研究罗利环素与人血清白蛋白的相互作用[J]. 光谱学与光谱分析, 2018, 38(3): 990. FANG Qing, DONG Cheng-yu, WANG Yu, LIU Ying. The Interaction between Rolitetracycline and Human Serum Albumin Using Multi-Spectral Methods and Molecular Modeling[J]. Spectroscopy and Spectral Analysis, 2018, 38(3): 990.

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