Effect of ponceau 4R on the spectral properties of trypsin
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摘要: 应用荧光光谱法、紫外可见吸收光谱法、傅立叶变换红外光谱和圆二色光谱法,研究了食用合成色素胭脂红与胰蛋白酶的结合性质,明确胭脂红与胰蛋白酶之间的结合常数、结合位点数（n）、作用力类型、结合距离（r）等信息。结果表明,胭脂红对胰蛋白酶内源荧光强度有较强的猝灭能力,荧光猝灭机制属于形成复合物的静态猝灭,由实验计算得出的热力学参数熵变和焓变均为负值,表明氢键和范德华力是驱动胭脂红-胰蛋白酶复合物形成的主要作用力,胭脂红与胰蛋白酶的结合常数达到104mol/L数量级,存在中等强度的亲和力。光谱分析表明胭脂红与胰蛋白酶结合引起胰蛋白酶的氨基酸残基微环境发生改变,胰蛋白酶的二级结构α-螺旋和无规则卷曲含量增加,β-折叠和β-转角的含量降低,多肽链有所收缩。Abstract: The binding characteristics between ponceau 4R and trypsin was investigated by fluorescence, UV-vis absorption, Fourier transform infrared ( FT-IR) and circular dichroism ( CD) spectroscopy, to obtain the binding constant, binding sites ( n) , the types of forces, the binding distance ( r) between ponceau 4R and trypsin, and other information. The results showed that ponceau 4R had a strong ability to quench the intrinsic fluorescence of trypsin, the quenching mechanism of trypsin by the ponceau 4R was a static process. The calculated thermodynamic parameters showed that the binding process was primarily driven by hydrogen bonds and van der Waals forces.The binding constant between them was 104 orders of magnitude, revealing that ponceau 4R can bind to trypsin with moderate affinity.Moreover, analysis of synchronous fluorescence, UV-vis absorption, Fourier transform infrared ( FT-IR) and CD spectra demonstrated that the binding interaction induced the microenvironment changes and conformational alteration of trypsin with increases in α-helix and random coil contents, and reduction of β-sheet and β-turn structures, resulting in partial shrinkage of the polypeptides of protein.
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Keywords:
- ponceau 4R /
- trypsin /
- fluorescence quenching /
- binding characteristic
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[1] Tabara A, Yamane C, Abe M, et al.Adsorption of xanthene food additive dyes to cellulose granules[J].Cellulose, 2011, 18 (1) :45-55.
[2] 曾盈, 蔡智鸣, 王振, 等.胭脂红对3T3细胞DNA损伤的SCGE检测[J].同济大学学报:医学版, 2008, 29 (1) :41-44. [3] 倪逸声, 张龙翔.胰蛋白酶工程研究进展[J].生物工程进展, 1992, 12 (1) :2-8. [4] 刘志栋.荧光法研究胭脂红, 柠檬黄及日落黄与BSA相互作用机理[D].哈尔滨:黑龙江大学, 2014. [5] 马明明, 同帜.几种光谱法研究牛血清白蛋白与胭脂红作用[J].光谱实验室, 2011, 28 (5) :2188-2193. [6] Mu Y, Lin J, Liu R.Interaction of sodium benzoate with trypsin by spectroscopic techniques[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2011, 83 (1) :130-135.
[7] Yan J, Zhang G, Hu Y, et al.Effect of luteolin on xanthine oxidase:Inhibition kinetics and interaction mechanism merging with docking simulation[J].Food Chemistry, 2013, 141 (4) :3766-3773.
[8] 丁琼, 倪永年.荧光光谱法研究丙烯酰胺与牛血清白蛋白的相互作用[J].南昌大学学报:理科版, 2012, 36 (6) :543-547. [9] 段晓燕.金纳米粒子猝灭Cd Te量子点荧光的研究[D].西安:陕西师范大学, 2013. [10] Wang Z, Li Y, Wang J, et al.Spectroscopic analyses on sonocatalytic damage to bovine serum albumin (BSA) induced by Zn O/hydroxylapatite (Zn O/HA) composite under ultrasonic irradiation[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2012, 94:228-234.
[11] Wu D, Yan J, Wang J, et al.Characterization of interaction between food colourant Allura Red AC and human serum albumin:multispectroscopic analyses and docking simulations[J].Food Chemistry, 2014, 170:423-429.
[12] Lakowicz J R, Weber G.Quenching of fluorescence by oxygen.Probe for structural fluctuations in macromolecules[J].Biochemistry, 1973, 12 (21) :4161-4170.
[13] Wu X, He W, Wang W, et al.Investigation of the interaction between (-) -epigallocatechin-3-gallate with trypsin andα-chymotrypsin[J].International Journal of Food Science&Technology, 2013, 48 (11) :2340-2347.
[14] 张秋兰, 倪永年.光谱和伏安法研究沙丁胺醇与牛血清白蛋白的相互作用[J].南昌大学学报:理科版, 2014, 38 (1) :69-73. [15] Wang R, Dou H, Yin Y, et al.Investigation of the interaction between isomeric derivatives and human serum albumin by fluorescence spectroscopy and molecular modeling[J].Journal of Luminescence, 2014, 154:8-14.
[16] Hu Y, Zhang G, Yan J.Detection of interaction between lysionotin and bovine serum albumin using spectroscopic techniques combined with molecular modeling[J].Molecular Biology Reports, 2014, 41 (3) :1693-1702.
[17] F9rster T, Sinanogly O.Modern quantum chemistry[M].New York:Academic Press, 1966.
[18] Xiang Y, Wu F.Study of the interaction between a new Schiff-base complex and bovine serum albumin by fluorescence spectroscopy[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2010, 77 (2) :430-436.
[19] Kalanur S S, Seetharamappa J, Kalalbandi V K A.Characterization of interaction and the effect of carbamazepine on the structure of human serum albumin[J].Journal of Pharmaceutical and Biomedical Analysis, 2010, 53 (3) :660-666.
[20] Ding F, Huang J L, Lin J, et al.A study of binding of C.1mordant red 3 with bovine serum albumin using fluorescence spectroscopy[J].Dyes and Pigments.2009, 82 (1) :65-70.
[21] Khodarahmi R, Karimi S A, Ashrafi Kooshk M R, et al.Comparative spectroscopic studies on drug binding characteristics and protein surface hydrophobicity of native and modified forms of bovine serum albumin:Possible relevance to change in protein structure/function upon non-enzymatic glycation[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2012, 89:177-186.
[22] Song W, Yu Z, Hu X, et al.Dissection of the binding of hydrogen peroxide to trypsin using spectroscopic methods and molecular modeling[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2014, 137:286-293.
[23] Kaur G, Tripathi S K.Investigation of trypsin-Cd Se quantum dot interactions via spectroscopic methods and effects on enzymatic activity[J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2015, 134:173-183.
[24] Zhang G W, Wang L, Pan J H.Probing the Binding of the Flavonoid Diosmetin to Human Serum Albumin by Multispectroscopic Techniques[J].Journal of Agricultural and Food Chemistry.2012, 60:2721-2729.
[25] Otsuka M, Fukui Y, Ozaki Y.Comparative evaluation of bioactivity of crystalline trypsin for drying by Fourier-transformed infrared spectroscopy[J].Colloids and Surfaces B:Biointerfaces, 2009, 69 (2) :194-200.
[26] Sarkar D, Mahata A, Das P, et al.Deciphering the perturbation of serum albumins by a ketocyanine dye:a spectroscopic approach[J].Journal of Photochemistry and Photobiology B:Biology, 2009, 96 (2) :136-143.
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