|
|
|
|
|
|
| Comparisons and Applications of Functional Equations for the Calculation of the Protein-Ligand Binding Constant Based on Fluorescence Spectral Data |
| ZHANG Jing1, GAO Xuan1, 2, JIN Liang1, WANG Hong-hui1, ZHOU Xi-ping1 |
1. Key Laboratory of Estuarine Ecological Security and Environmental Health, School of Environmental Science and Engineering, Xiamen University Tan Kah Kee College, Zhangzhou 363105, China
2. College of Environment and Ecology, Xiamen University, Xiamen 361102, China |
|
|
|
|
Abstract The binding interactions of proteins with ligands play important roles in the physiological and biochemical processes of organisms. Thus it is a research hotspot in many fields. A key objective of the investigations on Protein-Ligand binding is to obtain their binding constant (Kb) for evaluating the strength of their binding. Fluorescence spectroscopy is widely used in Protein-Ligand binding studies as its high sensitivity, convenience, rapidity and low cost. When applying fluorescence spectroscopy, it is a key step to obtain the Kb value through mathematical analysis of fluorescence data with the aid of a functional equation. However, due to the different application range of different equations, the Kb values obtained by different equations for the same system are often varied. This work discussed the derivation process and the application conditions of the functional equations for calculating the Kb value of Protein-Ligand binding using fluorescence spectral data. Based on this, we summarized the optimal equation for the calculation of the Kb value for Protein-Ligand binding modes of 1∶1 and 1∶n (n≥2) under different preconditions. The discussion suggested that the selection of the optimal equation is based on two preconditions: a: Whether the Protein-Ligand complex is fluorescent; b: Whether the ligand concentration is much higher than the protein concentration. On the other hand, the human serum albumin (HSA)-Norfloxacin (NFX) binding system was used as a model, and the difference of the Kb values obtained by the fitting of different functional equations was compared and the corresponding reasons were analyzed. The results showed that HSA bound with NFX in 1∶1 mode, and the Kb value was calculated to be 5.0×104 L·mol-1 at 298 K using the most appropriate equation (S12) for this system. However, the Kb values obtained by equation (S6) and (S24) were 28.8% and 48.6% higher than that by equation (S12) respectively, and the value obtained by equation (S17) was about two orders of magnitude higher. This intuitively showed the effect of choosing the inapplicable equation on the final result. In addition, the Kb value obtained by fluorescence spectroscopy in the present work was compared with those obtained by other techniques, indicating the relative reliability of fluorescence spectroscopy for the calculation of the Kb value in Protein-Ligand binding, and also pointing out the limitations of this method. This work warns us that when obtaining the Kb value of Protein-Ligand binding using fluorescence spectral data, it is very necessary to select the most suitable functional equation according to the preconditions (i.e. whether the hypothesis a and/or b are true or not), which will determine the reliability of the obtained results.
|
|
Received: 2019-11-14
Accepted: 2020-02-26
|
|
|
|
[1] Du X, Li Y, Xia Y L, et al. International Journal of Molecular Sciences, 2016, 17(2)：144.
[2] ZHAO Xiao-xue, GUO Ming, WANG Yan(赵晓雪，郭 明，王 燕). Spectroscopy and Spectral Analysis (光谱学与光谱分析), 2018, 38(11)：3420.
[3] Kairys V, Baranauskiene L, Kazlauskiene M, et al. Expert Opinion on Drug Discovery, 2019, 14(8)：755.
[4] Bakar, K A, Feroz, S R. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2019, 223：117337.
[5] Azimi O, Emami Z, Salari H, et al. Molecules, 2011, 16(12)：9792.
[6] Zhang L L, Liu Y C, Wang Y M. Food Chemistry, 2019, 301：125294.
[7] Rahman S, Rehman M T, Rabbani G, et al. International Journal of Molecular Sciences, 2019, 20(11)：2727.
[8] BI Shu-yun, DING Lan, SONG Da-qian, et al(毕淑云, 丁 兰, 宋大千,等). Acta Chimica Sinica(化学学报), 2005, 63(23): 2169.
[9] Zhang L W, Wang K, Zhang X X. Analytica Chimica Acta, 2007, 603(1)：101.
[10] Liu X, Song D Q, Zhang Q L, et al. Sensors and Actuators B-Chemical, 2006, 117(1)：188.
[11] Paul B K, Ghosh N, Mukherjee S. Journal of Physical Chemistry B, 2015, 119(41)：13093. |
| [1] |
YANG Jing1, LI Li1, LIANG Jian-dan1, HUANG Shan1, SU Wei1, WEI Ya-shu2, WEI Liang1*, XIAO Qi1*. Study on the Interaction Mechanism Between Thiosemicarbazide Aryl Ruthenium Complexes and Human Serum Albumin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(09): 2761-2767. |
| [2] |
YE Zi-yi, LIU Shuang, ZHANG Xin-feng*. Screening of DNA Dyes for Colorimetric Sensing Via Rapidly Inducing Gold Nanoparticles Aggregation[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(09): 2805-2810. |
| [3] |
REN Li-lei, PENG Yu-ling, WANG Shu-jun*, ZHANG Cheng-gen, CHEN Yu, WANG Xin-tong, MENG Xiao-ning. Fluorescence Spectroscopy for Studies on the Interaction Between Three Metalloporphyrins With Human Serum Albumin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2023, 43(03): 806-813. |
| [4] |
BAI Jun-peng1, 2, LI Bin1*, ZHANG Shu-juan2, CHEN Yi-mei1. Study on Norfloxacin Concentration Detection Based on Terahertz Time Domain Spectroscopy[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2021, 41(09): 2710-2716. |
| [5] |
HU Jing-jing, TONG Chang-lun*. Study on the Interaction Between Carbon Quantum Dots and Human Serum Albumin by Spectroscopic Methods[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2021, 41(04): 1107-1113. |
| [6] |
GUO Qing-ying1, LIU Min1,2*, ZHAO Yan-na2, WU Yu-shu2, SUN Bin2, LIU Jie1, HAN Jun2. Spectroscopic and Cytotoxicity Study on the Influence of (-)-Epigallocatechin-3-Gallate on the Interaction of Daunorubicin with Human Serum Albumin[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2020, 40(06): 1821-1827. |
| [7] |
HAN Zhong-bao1, WU Yu-hang1, MI Yuan-yuan1, LIU Li-yan1,2, SU Gui-tian1, YU Zhan1,2*. Isomeric Discrimination of Oleanolic and Ursolic Acids by Human Serum Albumin: a Joint Study of Fluorescence Spectroscopy and Molecular Docking[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2019, 39(07): 2190-2195. |
| [8] |
XIONG Shi-peng, CHEN Jian-bo*. Characterization of the Interactions between Alpha Arbutin and Human Serum Albumin with Spectroscopic Method and Molecular Docking[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2018, 38(11): 3489-3494. |
| [9] |
WANG A-mei1, TU Zong-cai1, 2, WANG Hui1*, MA Da1. Research in Glycated Progress and Products of Human Serum Albumin by Infrared and CD Spectroscopy[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2018, 38(10): 3090-3095. |
| [10] |
GONG Han-qing, CHEN Jian-bo*. The Interaction between 4-Ethyl-2-Methoxyphenol and Human Serum Albumin Studied by Spectroscopic and Molecular Docking Techniques[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2018, 38(06): 1869-1873. |
| [11] |
FANG Qing1, DONG Cheng-yu1, WANG Yu1, LIU Ying1,2*. The Interaction between Rolitetracycline and Human Serum Albumin Using Multi-Spectral Methods and Molecular Modeling[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2018, 38(03): 990-996. |
| [12] |
WANG Huan, GOU Xing-xing, PU Xiao-hua*, WANG Jiao, HU Xiao-bing, LI Zong-xiao. Exploration of the Interaction Mechanism between Doxorubicin Hydrochloride and DNA by Spectroscopic Techniques and Isothermal Titration Calorimetry[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2018, 38(02): 540-545. |
| [13] |
LIU Xing1, SHEN Peng1，2, LIANG An-wen1, YU Mei-lan1*. Study on Spectroscopy of Fullerols and Serum Albumin Interaction[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(11): 3530-3536. |
| [14] |
MA Xue-hong1, SONG Bai-ling2, XIAO Wen-jun2, GENG Jing2, LI Xin-xia1*, CHEN Jian3. Study on Interaction between Alliin and Bovine and Hunman Serum Albumin with Spectrometry[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(06): 1826-1830. |
| [15] |
LI Yuan, LI Kai, ZHAO Dong-e. Rapid Detection System for HSA Based on RRS[J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2017, 37(06): 1925-1929. |
|
|
|
|
