基于多光谱方法和计算模拟的细胞色素CYP1A2与阿奇霉素的结合机制研究

Study of the binding mechanism of cytochrome CYP1A2 to azithromycin based on multispectral approach and computational simulation

为了解阿奇霉素(AZI)在模拟的人体环境中与CYP1A2的结合情况,探索AZI在人体内的转化机制,本论文使用分子对接、分子动力学模拟、荧光光谱、紫外分光光度法、傅立叶变换红外光谱、圆二色光谱等多种实验方法,阐明AZI与CYP1A2之间的相互作用机制。分子对接结果表明AZI与CYP1A2的结合方式是半包裹并且以疏水作用力相结合。分子动力学模拟结果表明,CYP1A2-AZI复合物的均方根偏差(RMSD)值增大;均方根波动(RMSF)值表明复合物体系柔性较大;回旋半径(Rg)值表明蛋白质特定区域的结构松散。荧光猝灭光谱实验表明,CYP1A2与AZI是以静态猝灭机制结合在一起。热力学参数表明AZI与CYP1A2之间的主导作用力为疏水作用力。时间分辨光谱表明AZI对CYP1A2的机制为静态猝灭。紫外光谱实验表明AZI与CYP1A2反应生成了复合物。红外光谱实验结果表明CYP1A2的二级结构含量发生了改变。圆二色光谱证实AZI对CYP1A2的二级结构产生影响。

To understand the binding of azithromnycin(AZI)to CYP1A2 in a simulated human environment and to explore the con-version mechanism of azithromycin in human body.In this thesis,we used various experimental methods such as molecular docking.molecular dynamics simulation,fluorescence spectroscopy,UV spectrophotormetry,Fourier transform infrared spectroscopy and circu-lar dichroism spectroscopy to elucidate the interaction mechanism between AZI and CYP1A2.The molecular docking results showed that the binding mode of AZI and CYP1A2 was semi-ercapsulated and bound by hydrophobic force.Molecular dynamies simulationsshowed that the root mean square deviation(RMSD)values of the CYPIA2-AZ1 complexes incereased;the root mean square fluctua-tion(RMSF)values indicated that the complex systens were more flexible;the radius of gyration(Rg)values indicated that thestructure of specifie regions of the proteins was loose.Fluorescence bust spectroscopy experiments showed that CYPIA2 enzyme and AZI were bound together in a statie burst mechanism.Thermodynamic parameters indicate that the dominant force between AZI and CYP1A2 is a hydrophobic force.Time-resolved spectra indicatel that the mechanism of AZI on CYPIA2 was a static burst.UV spectroscopy experiments showed that AZI reacted with CYP1A2 to form a complex.Infrared spectroscopy experiments showed that the secondary structure content of CYP1A2 was altered.Circular dichroism spectroscopy confirmed the fft of AZI on the seconda-ry structure of CYP1A2.