光谱法研究盐酸非那吡啶与牛血清白蛋白的结合作用

运用光谱学方法研究了在生理pH值条件下盐酸非那吡啶(PHE)与牛血清白蛋白之间的结合作用.通过荧光光谱和紫外吸收光谱确定了盐酸非那吡啶对牛血清白蛋白的荧光猝灭机理.依据Scatchard方程测定了不同温度下该结合反应的结合常数和结合位点数.根据热力学方程讨论了两者间的主要作用力类型.结合同步荧光光谱分析了盐酸非那吡啶对牛血清白蛋白构象的影响.盐酸非那吡啶对牛血清白蛋白的荧光猝灭机制主要为静态猝灭和非辐射能量转移.在15,25,37℃时盐酸非那吡啶与牛血清白蛋白的结合常数Kb分别为2.47×107,9.15×106,4.36×106 L·mol-1,它们之间平均结合位点数n为1.结合反应的热力学参数为△H=-71.2 kJ·mol-1,△S=124.8 J·mol-1·K-1.热力学函数计算结果表明,该作用过程是一个熵增加,Gibbs自由能降低的自发分子间作用过程.依据F(o)rster能量转移理论确定PHE与BSA间的结合距离为1.61 nm.两者结合的主要作用力类型是静电作用力.盐酸非那吡啶在体内能够被血清蛋白存储和转运,但结合时对蛋白构象有一定影响.

Spectroscopic Studies on the Binding of Phenazopyridine Hydrochloride and Bovine Serum Albumin

The binding of phenazopyridine hydrochloride and bovine serum albumin under physiological conditions was studied by spectroscopic method. The quenching mechanism of the fluorescence of bovine serum albumin by phenazopyridine hydrochloride was studied with fluorescence and absorption spectroscopy. The binding constant Kb and the number of binding sites n were determined at different temperatures according to Scatchard equation, and the main binding force was discussed by thermodynamic equations. The effect of the drug on bovine serum albumin conformation was also studied by using synchronous fluorescence spectroscopy. The quenching mechanism of phenazopyridine hydrochloride to bovine serum albumin is static quenching and non-radiation energy transfer. The binding constants Kb at 15, 25 and 37 degrees C are 2.47 x 10(7), 9.15 x 10(6) and 4.36 x 10(6) mol(-1) with one binding site, respectively. The thermodynamic parameters of the reaction are DeltaH = -71.2 kJ x mol(-1), and DeltaS = 124.8 J x mol(-1) x K(-1). Binding phenazopyridine hydrochloride to bovine serum albumin is a spontaneous inter-molecular interaction in which entropy increases and Gibbs free energy decreases. The binding distance r between phenazopyridine hydrochloride and bovine serum albumin is 1.61 nm according to Forster theory of non-radiation energy transfer. The binding force is electrostatic interaction. Phenazopyridine hydrochloride can be deposited and transported by serum protein in vivo. Phenazopyridine hydrochloride does affect the serum protein conformation.