光谱和微量热法分析柑橘苷与BSA分子间作用及构建其理论模型
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摘要
光谱和微量热法分析柑橘苷(naringin,NAR)与牛血清白蛋白(bovine serum albumin,BSA)分子间作用,构建NAR与BSA分子间作用的理论模型。采用紫外-荧光光谱法解析Fōrster方程求得NAR与BSA分子间作用及分子间作用的临界距离r,等温滴定微量热技术测定NAR与BSA分子间作用的积分量热曲线,获得ΔH并通过Gibbs-Helmholtz方程获取ΔS和ΔG。基于光谱和微量热辅助分析,构建NAR与BSA分子间作用的理论模型。结果表明,光谱法测出NAR与BSA发生分子间作用,NAR与BSA分子间作用的临界距离为2.06 nm,表明NAR与BSA分子间作用为短程分子间作用。微量热法成功测定出NAR与BSA分子间的热力学参数ΔH<0,ΔS>0,ΔG<0,说明NAR与BSA分子间作用是自发进行的放热相互作用。依据Ross理论分析NAR与BSA分子间作用力主要是疏水作用力和静电作用力。模型构建结果说明,NAR与BSA分子间作用主要发生在BSA的domain IIA区域,NAR与BSA分子间作用力主要是静电作用力,兼有范德华作用力和氢键。实验与理论模型构建结果基本一致。本研究工作可为深入了解蛋白质与大分子化合物间的作用以及研究微观药理学机制提供有益的参考。
The intermolecular interaction between naringin( NAR) and bovine serum albumin( BSA)was studied by fluorescence spectrum and microcalorimetry. The theoretical model of the interaction between NAR and BSA was established. ΔH,ΔS and ΔG were obtained by using the isothermal titration microcalorimetric technique and Gibbs-Helmholtz equation. The Fōrster equation was used to analyze the critical distance between NAR and BSA in the UV-fluorescence spectra. Based on spectroscopy-and microcalorimetry-assisted analysis,the theoretical model of the interaction between NAR and BSA was established. The critical distance between NAR and BSA was 2. 06 nm,which indicated that the interaction between NAR and BSA was short-range intermolecular interaction. The results showed that thethermodynamic parameters ΔH < 0,ΔS > 0 and ΔG < 0 between NAR and BSA molecules have been successfully established by microcalorimetry. These microcalorimetry results showed that the interaction between NAR and BSA was spontaneous exothermic molecule interaction. According to the Ross theory analysis of NAR and BSA,intermolecular forces are mainly hydrophobic and electrostatic forces. Based on the analysis of UV-fluorescence spectra to obtain the binding distance,the intermolecular interactions between NAR and BSA were short-range. The theoretical model results showed that the intermolecular interaction of NAR and BSA occurred mainly in the domain IIA region of BSA. The NAR and BSA intermolecular forces are mainly electrostatic forces. Both van der Waals force and the presence of hydrogen bonds and the intermolecular interaction between BSA and NAR were a spontaneous process.The experimental results are basically consistent with the theoretical modeling results. These data can provide a useful reference for a comprehensive understanding of the intermolecular interactions between biological macromolecules and flavonoids and the study of microscopic pharmacological mechanisms.
The intermolecular interaction between naringin( NAR) and bovine serum albumin( BSA)was studied by fluorescence spectrum and microcalorimetry. The theoretical model of the interaction between NAR and BSA was established. ΔH,ΔS and ΔG were obtained by using the isothermal titration microcalorimetric technique and Gibbs-Helmholtz equation. The Fōrster equation was used to analyze the critical distance between NAR and BSA in the UV-fluorescence spectra. Based on spectroscopy-and microcalorimetry-assisted analysis,the theoretical model of the interaction between NAR and BSA was established. The critical distance between NAR and BSA was 2. 06 nm,which indicated that the interaction between NAR and BSA was short-range intermolecular interaction. The results showed that thethermodynamic parameters ΔH < 0,ΔS > 0 and ΔG < 0 between NAR and BSA molecules have been successfully established by microcalorimetry. These microcalorimetry results showed that the interaction between NAR and BSA was spontaneous exothermic molecule interaction. According to the Ross theory analysis of NAR and BSA,intermolecular forces are mainly hydrophobic and electrostatic forces. Based on the analysis of UV-fluorescence spectra to obtain the binding distance,the intermolecular interactions between NAR and BSA were short-range. The theoretical model results showed that the intermolecular interaction of NAR and BSA occurred mainly in the domain IIA region of BSA. The NAR and BSA intermolecular forces are mainly electrostatic forces. Both van der Waals force and the presence of hydrogen bonds and the intermolecular interaction between BSA and NAR were a spontaneous process.The experimental results are basically consistent with the theoretical modeling results. These data can provide a useful reference for a comprehensive understanding of the intermolecular interactions between biological macromolecules and flavonoids and the study of microscopic pharmacological mechanisms.
引文
[1]Lagazzo A,Barberis F,Carbone C,et al.Molecular level interactions in brushite-aminoacids composites[J].Mater Sci Eng C Mater Biol Appl,2017,70(Pt 1):721-727
[2]Bjrklund G,Aaseth J,Ajsuvakova OP,et al.Molecular interaction between mercury and selenium in neurotoxicity[J].Coordin Chem Rev,2016,332:30-37
[3]Ullah H.Inter-molecular interaction in polypyrrole/Tio2:A DFT study[J].J Alloys Compounds,2017,692:140-148
[4]Han XL,Tian FF,Ge YS,et al.Spectroscopic,structural and thermodynamic properties of chlorpyrifos bound to serum albumin:A comparative study between BSA and HSA[J].J Photochem Photobiol B,2012,109:1-11
[5]Liu Y,Cao ZZ,Wang J,et al.The interaction mechanism between anionic or cationic surfactant with HSA by using spectroscopy,calorimetry and molecular docking methods[J].J Mol Liquids,2016,224:1008-1015
[6]顾奕,郭明,吕达,等.毛细管电泳法考察全氟辛酸与人血清白蛋白的相互作用.色谱(Gu Y,Guo M,lv D,et al.Investigation on the interaction between pentadecafluorooctanoic acid and human serum albumin by capillary electrophoresis[J].Chin J Chromatogr),2018,36(1):69-77
[7]Ramakrishnan M,Alves De Melo F,Kinsey BM,et al.Probing cocaine-antibody interactions in buffer and human serum[J].PLo S One,2012,7(7):e40518
[8]李伊培,高丽华,张连成,等.重组人VEGF可溶性受体的表达纯化及结合性质初探[J].生物技术通讯(Li YP,Gao LH,Zhang LC,et al.Expression and purification of recombinant human soluble VEGF receptor and preliminary study of its binding affinity[J].Lett Biotechnol),2014,25(2):165-169
[9]Liu Y,Liu R.Spectroscope and molecular model identify the behavior of doxorubicin-SPION binding to bovine hemoglobin[J].Int J Biol Macromol,2015,79:564-569
[10]Jing M,Song W,Liu R.Binding of copper to lysozyme:Spectroscopic,isothermal titration calorimetry and molecular docking studies[J].Spectrochim Acta A Mol Biomol Spectrosc,2016,164:103-109
[11]Shi JH,Pan DQ,Jiang M,et al.Binding interaction of ramipril with bovine serum albumin(BSA):Insights from multispectroscopy and molecular docking methods[J].J Photochem Photobiol B,2016,164:103-111
[12]Guo M,Lu X,Wang Y,et al.Comparison of the interaction between lactoferrin and isomeric drugs[J].Spectrochim Acta A Mol Biomol Spectrosc,2017,173:593-607
[13]何文英,姚小军,刘鹏军,等.分子模拟与光谱法研究愈创木酚和人免疫球蛋白的键合[J].中国科学B辑(He WY,Yao XJ,Liu PJ,et al.olecular modeling and spectroscopic studies on the binding of guaiacol and human immunoglobulin[J].Sci China Ser B),2007,37(1):54-63
[14]王燕,郭明,王晓萌.厄他培南与人血清清蛋白体外相互作用的理化特性[J].中国生物化学与分子生物学报(Wang Y,Guo M,Wang XM.Physical and chemical characteristics of the in vitro interaction of Ertapenem with human serum albumin[J].Chin J Biochem Mol Biol),2016,32(8):892-900
[15]Ross PD,Subramanian S.Thermodynamics of protein association reactions:forces contributing to stability[J].Biochemistry,1981,20(11):3096-3102
[16]杨荣荣.应用光谱法及分子对接研究黄酬类化合物与牛血清白蛋白的结合特性[D].沈阳:大连工业大学(Yang RR.Study on binding characteristics of flavonoids with BSA by spectroscopic and molecular docking methods[D].Dalian Polytechnic University,Shenyang),2014
[17]Roy AS,Tripathy DR,Chatterjee A,et al.A spectroscopic study of the interaction of the antioxidant naringin with bovine serum albumin[J].J Biophys Chem,2010,1(3):141-152
[18]Ma R,Pan H,Shen T,et al.Interaction of flavonoids from woodwardia unigemmata with bovine serum albumin(BSA):application of spectroscopic techniques and molecular modeling methods[J].Molecules,2017,22(8):1317
[19]崔艳花,郭春梅,孙明忠,等.系列黄酮化合物与血清白蛋白结合及其结构相关性[J].中国生化药物杂志(Cui YH,Guo CM,Sun MZ,et al.The binding and structural association of flavonoids with bovine serum albumin[J].Chin J Biochem Pharmaceutics),2012,33(6):728-731
[20]王永杰,赵东霞,杨忠志.乙酰胆碱酯酶与抑制剂分子对接的ABRRMσπ/MM研究[J].分子科学学报(Wang YP,Zhao DX,Yang ZZ.Study on interaction of ACh E and its ligands by ABEEMσπ/MM method[J].J Mol Sci),2012,28(2):115-122
[21]Sivashanmugam M,Raghunath C,Vetrivel U.Virtual screening studies reveal linarin as a potential natural inhibitor targeting CDK4 in retinoblastoma[J].J Pharmacol Pharmacotherapeutics,2013,4(4):256-264
[22]Ali MS,AI-Lohedan HA.Deciphering the interaction of procaine with bovine serum albumin and elucidation of binding site:A multi spectroscopic and molecular docking study[J].J Mol Liquids.2017,236:232-240
[23]赵琦.基于分子对接技术的小分子-蛋白质相互作用研究[D].无锡:江南大学(Zhao Q.The research on ligand-protein interaction using molecular docking[D].Jiangnan University,Wuxi),2009
[24]Zhuang S,Wang H,Ding K,et al.Interactions of benzotriazole UV stabilizers with human serum albumin:Atomic insights revealed by biosensors,spectroscopies and molecular dynamics simulations[J].Chemosphere,2016,144:1050-1059
[25]Forli S,Olson AJ.A force field with discrete displaceable waters and desolvations entropy for hydrated ligand docking[J].J Med Chem,2012,55(2):623-638
[26]Morris GM,Goodsell DS,Halliday RS,et al.Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function[J].J Comput Chem,2015,19(14):1639-1662
[2]Bjrklund G,Aaseth J,Ajsuvakova OP,et al.Molecular interaction between mercury and selenium in neurotoxicity[J].Coordin Chem Rev,2016,332:30-37
[3]Ullah H.Inter-molecular interaction in polypyrrole/Tio2:A DFT study[J].J Alloys Compounds,2017,692:140-148
[4]Han XL,Tian FF,Ge YS,et al.Spectroscopic,structural and thermodynamic properties of chlorpyrifos bound to serum albumin:A comparative study between BSA and HSA[J].J Photochem Photobiol B,2012,109:1-11
[5]Liu Y,Cao ZZ,Wang J,et al.The interaction mechanism between anionic or cationic surfactant with HSA by using spectroscopy,calorimetry and molecular docking methods[J].J Mol Liquids,2016,224:1008-1015
[6]顾奕,郭明,吕达,等.毛细管电泳法考察全氟辛酸与人血清白蛋白的相互作用.色谱(Gu Y,Guo M,lv D,et al.Investigation on the interaction between pentadecafluorooctanoic acid and human serum albumin by capillary electrophoresis[J].Chin J Chromatogr),2018,36(1):69-77
[7]Ramakrishnan M,Alves De Melo F,Kinsey BM,et al.Probing cocaine-antibody interactions in buffer and human serum[J].PLo S One,2012,7(7):e40518
[8]李伊培,高丽华,张连成,等.重组人VEGF可溶性受体的表达纯化及结合性质初探[J].生物技术通讯(Li YP,Gao LH,Zhang LC,et al.Expression and purification of recombinant human soluble VEGF receptor and preliminary study of its binding affinity[J].Lett Biotechnol),2014,25(2):165-169
[9]Liu Y,Liu R.Spectroscope and molecular model identify the behavior of doxorubicin-SPION binding to bovine hemoglobin[J].Int J Biol Macromol,2015,79:564-569
[10]Jing M,Song W,Liu R.Binding of copper to lysozyme:Spectroscopic,isothermal titration calorimetry and molecular docking studies[J].Spectrochim Acta A Mol Biomol Spectrosc,2016,164:103-109
[11]Shi JH,Pan DQ,Jiang M,et al.Binding interaction of ramipril with bovine serum albumin(BSA):Insights from multispectroscopy and molecular docking methods[J].J Photochem Photobiol B,2016,164:103-111
[12]Guo M,Lu X,Wang Y,et al.Comparison of the interaction between lactoferrin and isomeric drugs[J].Spectrochim Acta A Mol Biomol Spectrosc,2017,173:593-607
[13]何文英,姚小军,刘鹏军,等.分子模拟与光谱法研究愈创木酚和人免疫球蛋白的键合[J].中国科学B辑(He WY,Yao XJ,Liu PJ,et al.olecular modeling and spectroscopic studies on the binding of guaiacol and human immunoglobulin[J].Sci China Ser B),2007,37(1):54-63
[14]王燕,郭明,王晓萌.厄他培南与人血清清蛋白体外相互作用的理化特性[J].中国生物化学与分子生物学报(Wang Y,Guo M,Wang XM.Physical and chemical characteristics of the in vitro interaction of Ertapenem with human serum albumin[J].Chin J Biochem Mol Biol),2016,32(8):892-900
[15]Ross PD,Subramanian S.Thermodynamics of protein association reactions:forces contributing to stability[J].Biochemistry,1981,20(11):3096-3102
[16]杨荣荣.应用光谱法及分子对接研究黄酬类化合物与牛血清白蛋白的结合特性[D].沈阳:大连工业大学(Yang RR.Study on binding characteristics of flavonoids with BSA by spectroscopic and molecular docking methods[D].Dalian Polytechnic University,Shenyang),2014
[17]Roy AS,Tripathy DR,Chatterjee A,et al.A spectroscopic study of the interaction of the antioxidant naringin with bovine serum albumin[J].J Biophys Chem,2010,1(3):141-152
[18]Ma R,Pan H,Shen T,et al.Interaction of flavonoids from woodwardia unigemmata with bovine serum albumin(BSA):application of spectroscopic techniques and molecular modeling methods[J].Molecules,2017,22(8):1317
[19]崔艳花,郭春梅,孙明忠,等.系列黄酮化合物与血清白蛋白结合及其结构相关性[J].中国生化药物杂志(Cui YH,Guo CM,Sun MZ,et al.The binding and structural association of flavonoids with bovine serum albumin[J].Chin J Biochem Pharmaceutics),2012,33(6):728-731
[20]王永杰,赵东霞,杨忠志.乙酰胆碱酯酶与抑制剂分子对接的ABRRMσπ/MM研究[J].分子科学学报(Wang YP,Zhao DX,Yang ZZ.Study on interaction of ACh E and its ligands by ABEEMσπ/MM method[J].J Mol Sci),2012,28(2):115-122
[21]Sivashanmugam M,Raghunath C,Vetrivel U.Virtual screening studies reveal linarin as a potential natural inhibitor targeting CDK4 in retinoblastoma[J].J Pharmacol Pharmacotherapeutics,2013,4(4):256-264
[22]Ali MS,AI-Lohedan HA.Deciphering the interaction of procaine with bovine serum albumin and elucidation of binding site:A multi spectroscopic and molecular docking study[J].J Mol Liquids.2017,236:232-240
[23]赵琦.基于分子对接技术的小分子-蛋白质相互作用研究[D].无锡:江南大学(Zhao Q.The research on ligand-protein interaction using molecular docking[D].Jiangnan University,Wuxi),2009
[24]Zhuang S,Wang H,Ding K,et al.Interactions of benzotriazole UV stabilizers with human serum albumin:Atomic insights revealed by biosensors,spectroscopies and molecular dynamics simulations[J].Chemosphere,2016,144:1050-1059
[25]Forli S,Olson AJ.A force field with discrete displaceable waters and desolvations entropy for hydrated ligand docking[J].J Med Chem,2012,55(2):623-638
[26]Morris GM,Goodsell DS,Halliday RS,et al.Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function[J].J Comput Chem,2015,19(14):1639-1662