Gokavi Naveen M, Nandibewoor Sharanappa T, Gowda Jayant I
P. G. Department of Studies in Chemistry, Karnatak University, Dharwad, Karnataka, 580003, India.
Department of Chemistry, Karnatak Science College, Dharwad, Karnataka, 580001, India.
J Fluoresc. 2023 Sep;33(5):2061-2073. doi: 10.1007/s10895-023-03199-y. Epub 2023 Mar 28.
The interaction of orphenadrine hydrochloride (ORD) with the model protein, bovine serum albumin (BSA), was investigated using a variety of spectroscopic techniques such as steady-state fluorescence, ultraviolet-visible, Fourier transform infrared, 3-D spectroscopy, and electrochemical methods under physiological conditions. Stern-Volmer plots were used to calculate fluorescence quenching at various temperatures. The findings point to a static quenching mechanism between ORD and BSA. At various reaction times, the binding sites (n) and binding constants (K) of ORD to BSA were recorded. Thermodynamic parameters ∆H, ∆S and ∆G between ORD and BSA were calculated and reported. The average binding distance (r) between the donor (BSA) and acceptor (ORD) molecules was predicted using Förster's theory. Three-dimensional fluorescence spectra, Fourier transform infrared spectra, and synchronous fluorescence studies all supported the alternations in protein structure following the interaction with ORD. A displacement study using site probes such as warfarin, ibuprofen, and digitoxin confirmed ORD binding at Sudlow's site I of BSA. The effect of common metal ions such as Cu, Ni, Ca, Co, and Zn on binding constant values was investigated and reported.
在生理条件下，使用多种光谱技术，如稳态荧光、紫外可见光谱、傅里叶变换红外光谱、三维光谱和电化学方法，研究了盐酸奥芬那君（ORD）与模型蛋白牛血清白蛋白（BSA）的相互作用。利用斯特恩-沃尔默图计算不同温度下的荧光猝灭。研究结果表明ORD与BSA之间存在静态猝灭机制。记录了不同反应时间下ORD与BSA的结合位点（n）和结合常数（K）。计算并报告了ORD与BSA之间的热力学参数∆H、∆S和∆G。利用福斯特理论预测了供体（BSA）和受体（ORD）分子之间的平均结合距离（r）。三维荧光光谱、傅里叶变换红外光谱和同步荧光研究均支持与ORD相互作用后蛋白质结构的变化。使用华法林、布洛芬和地高辛等位点探针进行的置换研究证实了ORD在BSA的Sudlow位点I处的结合。研究并报告了常见金属离子如Cu、Ni、Ca、Co和Zn对结合常数的影响。
