【摘要】 以牛血清白蛋白-TritonX-100和牛血清白蛋白-盐酸西布曲明两个体系为实例,考察了以血清白蛋白和小分子化合物为荧光检测对象所获得的相互作用信息的差异。发现两种方法获得的分子间结合常数差异显著,说明在以血清白蛋白为检测对象的传统荧光光谱法中,以色氨酸基团的荧光光谱所表达的信息来代表整个蛋白分子的相互作用信息是不准确的。文章提出了以荧光小分子化合物为检测对象的改进荧光光谱方法和荧光背景扣除方法,前者能全面表达相互作用过程中分子的整体信息,后者实现了荧光光谱交叠体系的荧光光谱法相互作用分析。更多还原

【Abstract】 The fluorescence spectroscopy is widely used in the analysis of the interaction between serum albumin (SA) and micromolecule compound, in which SA is measured as the fluorescence source to provide interaction information. A modified method has been compared with the classical method in the present paper, where the micromolecule compounds were used as the interested analytes whose fluorescence spectra were measured. The binding constants obtained based on the classical and modified methods were found much different by studying the two systems, BSA-TX and BSA-SH. The result shows that the classical method has the drawback of taking the fluorescence source of tryptophan instead of all other amino acids groups in SA molecule. Two novel methods were then reported, namely the modified fluorescence spectrometry method and fluorescence background subtracting method. The former could reflect the whole information of the interaction system, while the latter effectively overcame the overlay of spectra in the system, where the fluorescence spectra of the two components interfere with each other terribly. The effectivity of the two novel methods has been proved by ultra-violet spectrometry.更多还原