离子淌度质谱结合荧光光谱法研究5-氟尿苷与铜锌超氧化物歧化酶的相互作用

摘要

分别利用离子淌度质谱和荧光光谱方法研究了5-氟尿苷(5-Furd)与铜锌超氧化物歧化酶(SOD1)的相互作用.离子淌度质谱研究结果表明, 5-Furd能够稳定SOD1构型, 减缓其碰撞诱导去折叠;外源荧光光谱方法进一步证明5-Furd可以抑制SOD1的聚集.利用荧光光谱研究了不同温度下5-Furd与SOD1的相互作用, 阐明了二者的作用机制、 结合常数和结合位点数, 并根据热力学函数推断二者的作用力类型为典型的疏水作用.%The interaction between 5-Furd and Cu, Zn superoxide dismutase(SOD1) was investigated by means of electrospray-ion mobility mass spectrometry(ESI-IMS-MS) and fluorescence spectroscopy.The result of ESI-IMS-MS showed that 5-Furdis could stabilize the conformation of SOD1 against unfolding transition at different collision voltages.Through the ThT fluorescence method, it was further testified that 5-Furd could {inhibit} the aggregation of apo-SOD1.In addition, the interaction between SOD1 and 5-Furd at different temperatures was investigated by fluorescence spectrometry, and the characters of their interaction were elucidated including the interaction mechanism, the binding constant, binding site number and the corresponding thermo-dynamic parameters(ΔH, ΔS and ΔG).According to the thermo-dynamic parameters of interaction, it was found that the hydrophobic forces play an important role on their interaction.