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研究新型碳青霉烯类抗菌素厄他培南(ertapenem,ERT)与人血清清蛋白(human serum albumin,HSA)的体外相互作用的物理化学特性。模拟生理条件下,计算机模拟技术结合荧光光谱和紫外光谱,研究ERT与HSA相互作用机制,荧光光谱实验中,Kq值远大于2.0×1010L·(mol·s)-1,ERT对HSA荧光猝灭的Stern-Volmer曲线有良好的线性关系,表明ERT与HSA的相互作用表现为静态结合过程。HSA的最大发射波长发生轻微红移,说明HSA的微环境发生了改变。ERT与HSA的分子结合距离r值较小,说明发生能量转移现象。同步荧光技术解析出ERT对HSA的结构域微区构象产生影响,使色氨酸残基周围的微区构象及结合位域的疏水性发生改变。荧光相图技术解析出ERT与HSA相互反应呈线性,说明HSA构象型态的变迁为“二态”模型。HSA与ERT相互作用的热力学参数及分子模拟技术建立ERT-HSA结合模型,表明ERT与HSA的相互作用力主要是疏水作用力,兼有氢键作用力的存在。荧光偏振定量证明,HSA与ERT相互作用过程中生成了非共价复合物。光谱实验与计算机模拟结果基本一致,其结果可为研究ERT与HSA相互作用本质提供一定参考。
To study the physico-chemical properties of the in vitro interaction between a novel carbapenem antibiotic ertapenem (ERT) and human serum albumin (HSA). Under simulated physiological conditions, the interaction between ERT and HSA was studied by computer simulation with fluorescence and UV spectroscopy. The Kq value was much higher than 2.0 × 1010 L · (mol · s) -1 in fluorescence spectroscopy. The fluorescence quenching The Stern-Volmer curve showed a good linear relationship, indicating that the interaction between ERT and HSA showed a static binding process. A slight redshift of the maximum emission wavelength of HSA indicates that the microenvironment of HSA has changed. ERT and HSA molecular binding distance r value is small, indicating the occurrence of energy transfer phenomenon. Synchronous fluorescence analysis of ERT on the HSA domain microstructure conformation impact, the tryptophan residues around the micro-area conformation and the binding domain of hydrophobic change. Fluorescence phase diagram analysis of linear response of ERT and HSA was linear, indicating the conformational changes of HSA as “two state ” model. The thermodynamic parameters and molecular modeling techniques for the interaction between HSA and ERT were established ERT-HSA binding model, indicating that the interaction between ERT and HSA is mainly the hydrophobic force, both hydrogen bonds exist. Fluorescence polarization quantification demonstrated that a non-covalent complex was formed during the interaction of HSA with ERT. The results of spectral experiments and computer simulation are basically the same, and the results can provide certain reference for studying the nature of the interaction between ERT and HSA.