光谱法研究丙烯酰胺与牛血清白蛋白的相互作用

A STUDY OF THE INTERACTION BETWEEN ACRYLAMIDE AND BOVINE SERUM ALBUMIN USING SPECTROSCOPY METHODS

本文从光谱学的角度研究了有毒物质丙烯酰胺(AA)与牛血清白蛋白(BSA)间的相互作用。实验结果表明,在pH=7.0和室温条件下,AA可通过静态猝灭的方式有效地猝灭BSA的荧光,AA的加入影响了BSA的构象。通过荧光数据计算可得到AA与BSA的结合位点数(n=0.933)及表观绑定常数(KA=5.55×103mol.kg-1,由此表明AA与BSA的作用中存在单独的结合位点。实验证明最佳实验条件是,AA与BSA反应的时间为50 m in,离子强度为0.05 mol.kg-1,pH值为7.0。本文还利用共振散射光谱法、紫外可见吸收光谱法和傅里叶红外光谱法证实了AA对BSA二级结构的影响。AA与BSA相互作用的研究对生物分子的毒理性研究有重要作用。

The interaction between acrylamide（AA） and bovine serum albumin（BSA） was studied mainly from a spectroscopic angle.Under pH 7.0,AA effectively quenched the intrinsic fluorescence of BSA via static quenching and conformational changes of BSA were observed.The number of binding sites（n = 0.933） and apparent binding constant（KA = 5.55 ×103 M-1） were calculated by relevant fluorescence data,and thus indicated the existence of just a single binding site in BSA for AA.According to the controlled experiments,the optimal reaction conditions were obtained as follows： the reaction time of 50 min,ionic strength of 0.05 mol·kg-1,and pH = 7.0.The effect of AA on the conformation of BSA can also be deduced by means of resonance scattering spectrum,UV-visible absorption spectrum and FT-IR measurement.Therefore,The binding study of AA with BSA is of great importance in the research of the effect of toxicity to biomolecules.