College of Biology and Food Engineering, Guangdong University of Petrochemical Technology, No. 1, Kechuang Road, Maonan District, Maoming 525000, China.
College of Animal Science and Technology, Yangtze University, 88 Jingmi Road, Jingzhou District, Jingzhou 434025, China.
Molecules. 2024 Jan 30;29(3):655. doi: 10.3390/molecules29030655.
The interaction between human serum albumin (HSA) and hispidin, a polyketide abundantly present in both edible and therapeutic mushrooms, was explored through multispectral methods, hydrophobic probe assays, location competition trials, and molecular docking simulations. The results of fluorescence quenching analysis showed that hispidin quenched the fluorescence of HSA by binding to it via a static mechanism. The binding of hispidin and HSA was validated further by synchronous fluorescence, three-dimensional fluorescence, and UV/vis spectroscopy analysis. The apparent binding constant (Ka) at different temperatures, the binding site number (n), the quenching constants (Ksv), the dimolecular quenching rate constants (Kq), and the thermodynamic parameters (∆G, ∆H, and ∆S) were calculated. Among these parameters, ∆H and ∆S were determined to be 98.75 kJ/mol and 426.29 J/(mol·K), respectively, both exhibiting positive values. This observation suggested a predominant contribution of hydrophobic forces in the interaction between hispidin and HSA. By employing detergents (SDS and urea) and hydrophobic probes (ANS), it became feasible to quantify alterations in Ka and surface hydrophobicity, respectively. These measurements confirmed the pivotal role of hydrophobic forces in steering the interaction between hispidin and HSA. Site competition experiments showed that there was an interaction between hispidin and HSA molecules at site I, which situates the IIA domains of HSA, which was further confirmed by the molecular docking simulation.
通过多光谱方法、疏水性探针实验、定位竞争实验和分子对接模拟研究了人血清白蛋白（HSA）与广泛存在于食用和药用蘑菇中的聚酮化合物毛喉素之间的相互作用。荧光猝灭分析的结果表明，毛喉素通过静态机制与 HSA 结合而猝灭其荧光。同步荧光、三维荧光和紫外/可见光谱分析进一步验证了毛喉素与 HSA 的结合。在不同温度下计算了表观结合常数（Ka）、结合位点数（n）、荧光猝灭常数（Ksv）、双分子猝灭速率常数（Kq）和热力学参数（∆G、∆H 和 ∆S）。这些参数中，∆H 和 ∆S 分别为 98.75 kJ/mol 和 426.29 J/（mol·K），均为正值。这表明在毛喉素与 HSA 的相互作用中主要存在疏水作用力。通过使用去污剂（SDS 和尿素）和疏水性探针（ANS），分别可以定量测定 Ka 和表面疏水性的变化。这些测量证实了疏水作用力在引导毛喉素与 HSA 相互作用中的关键作用。位点竞争实验表明，毛喉素与 HSA 分子在 HSA 的 IIA 结构域的位点 I 处存在相互作用，分子对接模拟进一步证实了这一点。
