分类： 物理学 >> 核物理学 提交时间： 2023-06-18 合作期刊: 《Nuclear Science and Techniques》
MA Yu-Xing
ZHONG Rui-Bo
GUO Jun
LIU Yu-Shuang
YUAN Ming
BAI Zhi-Jun
LIU Tao-Tao
ZHAO Xin-Min
ZHANG Feng
摘要： Strategies for labeling proteins with fluorophores are always important for biotechnology. Here we take a model protein (bovine serum albumin) and a typical fluorophore (rhodamine B) to demonstrate a direct labeling method just by physical adsorption. In combination with size exclusion chromatography and the Scartchard equation, we have developed a facile analysis method for calculating the binding constant and binding sites. The molecular docking method has been used to study the binding site in amino acid level.
分类： 物理学 >> 核物理学 提交时间： 2023-06-18 合作期刊: 《Nuclear Science and Techniques》
摘要： Fluorescence spectroscopy was employed to investigate the interaction between fluorophore fluoresceinamine (FA) and bovine serum albumin (BSA) under physiological conditions. In the mechanism discussion, it was proved that the fluorescence quenching of BSA by FA is a result of the formation of a BSA-FA complex. Fluorescence quenching constants were determined using the modified Stern-Volmer equation to provide a measure of the binding affinity between FA and BSA. The results of the thermodynamic parameters G, H, and S at different temperatures indicated that several kinds of interactions, except for the electrostatic interactions play cooperative roles in BSA-FA association. Furthermore, the conformation of BSA upon interaction with FA was also studied by synchrotron fluorescence spectroscopy.
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