Study on Interaction between Coomassie Brilliant Blue G-250 and Bovine Serum Albumin by Fluorescence Spectroscopy and Molecular Modeling
WANG Yong-gang1, YANG Guang-rui1, MA Xue-qing2, LENG Fei-fan1, MA Jian-zhong1*, WANG Xiao-li3
1. School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China
2. State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
3. Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agricultural Sciences, Lanzhou 730050, China
Abstract：The interaction of coomassie brilliant blue G-250 (CBBG-250) with bovine serum albumin (BSA) was investigated by the methods of fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR) and circular dichroism (CD). Fluorescence data showed that the quenching of BSA by CBBG-250 was result of forming the complex of BSA-CBBG-250, and CBBG-250 effectively quenched the intrinsic fluorescence of BSA via static quenching. According to Stern-Volmer equation, the binding parameters between CBBG-250 and BSA were determined. The enthalpy change (ΔH) and entropy change (ΔS) were calculated to be -4.38 kJ·mol-1 and -6.16 J·mol-1·K-1, indicating that the hydrogen bonds and hydrophobic interactions played a dominant role in the binding. The conformational investigation revealed the α-helical structure was decreased and the polypeptides of BSA were slightly folded with the addition of CBBG-250 by synchronous fluorescence. Fourier infrared spectrum showed that the peak position of amide band Ⅰ (1 600～1 700 cm-1) and amide band Ⅱ (1 600～1 500 cm-1) of the BSA characteristic absorbing peaks was blueshifted. The amide band I moved from 1 650 to 1 710 cm-1, and amide band Ⅱ moved from 1 573 to 1 544 cm-1, which indicated the structure of alpha helix features (1 650～1 658 cm-1) and beta-fold (1 620～1 640 cm-1 and 1 645 cm-1) had changed. The content of α-helical structure decreased from 42.15% to 1.27% by circular dichroism. The main reasons were the microenvironment polarity of tryptophan changed with the addition of CBBG-250 increased, two interaction between BSA and CBBG-250 including the hydrogen bonding and van der Waals force occurred, and then the secondary structure of BSA was changed, which was confirmed by Molecular modeling.
王永刚，杨光瑞，马雪青，冷非凡，马建忠，王晓力. 荧光光谱法和分子模拟技术研究考马斯亮蓝G-250与牛血清白蛋白的相互作用[J]. 光谱学与光谱分析, 2017, 37(08): 2474-2479.
WANG Yong-gang, YANG Guang-rui, MA Xue-qing, LENG Fei-fan, MA Jian-zhong, WANG Xiao-li. Study on Interaction between Coomassie Brilliant Blue G-250 and Bovine Serum Albumin by Fluorescence Spectroscopy and Molecular Modeling. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(08): 2474-2479.
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