2种天然抗氧化剂与鲢鱼肌球蛋白的相互作用
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摘要
研究2种天然抗氧化剂表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)、白藜芦醇(resveratrol,RE)与鲢鱼肌球蛋白之间的相互作用。采用荧光光谱与圆二色谱技术分别探究EGCG、RE与鲢鱼肌球蛋白分子间相互作用的结合情况和其对肌球蛋白微观结构的影响。荧光光谱结果表明:EGCG、RE与鲢鱼肌球蛋白之间发生相互作用均会导致肌球蛋白荧光猝灭现象的发生,且荧光猝灭方式都为静态猝灭。通过热力学数据分析得出EGCG、RE与肌球蛋白分子结合的主要作用力是氢键和范德华力;并利用Stern-Volmer方程处理数据得到EGCG、RE与肌球蛋白在不同温度条件下的结合常数和结合位点数。圆二色谱及表面疏水性结果表明,EGCG、RE诱导了肌球蛋白结构的变化,EGCG使肌球蛋白α-螺旋相对含量增加,表面疏水性降低;而RE对肌球蛋白二级结构无明显作用,但会使其表面疏水性增加。
In this investigation, the interaction between two natural antioxidants epigallocatechin gallate(EGCG), resveratrol(RE) and silver carp myosin was studied. The changes in the intermolecular interaction and microstructure of myosin were evaluated by fluorescence and circular dichroism spectroscopy. The results showed that the interaction of silver carp myosin with EGCG or RE led to fluorescence quenching of silver carp myosin in a static quenching mode. Thermodynamic analysis showed that the main driving forces for EGCG and RE binding to myosin molecules were hydrogen bonding and van der Waals force. The Stern-Volmer equation was used to obtain the binding constants and the number of binding sites at different temperatures. Circular dichroism(CD) spectroscopy and surface hydrophobicity analysis showed that EGCG and RE induced changes in myosin structure; EGCG increased the content of α-helix and decreased the surface hydrophobicity,while RE had no effect on the secondary structures of myosin and increased the surface hydrophobicity.
In this investigation, the interaction between two natural antioxidants epigallocatechin gallate(EGCG), resveratrol(RE) and silver carp myosin was studied. The changes in the intermolecular interaction and microstructure of myosin were evaluated by fluorescence and circular dichroism spectroscopy. The results showed that the interaction of silver carp myosin with EGCG or RE led to fluorescence quenching of silver carp myosin in a static quenching mode. Thermodynamic analysis showed that the main driving forces for EGCG and RE binding to myosin molecules were hydrogen bonding and van der Waals force. The Stern-Volmer equation was used to obtain the binding constants and the number of binding sites at different temperatures. Circular dichroism(CD) spectroscopy and surface hydrophobicity analysis showed that EGCG and RE induced changes in myosin structure; EGCG increased the content of α-helix and decreased the surface hydrophobicity,while RE had no effect on the secondary structures of myosin and increased the surface hydrophobicity.
引文
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[29]尹燕霞,向本琼,佟丽.荧光光谱法在蛋白质研究中的应用[J].实验技术与管理,2010,27(2):33-36;40.DOI:10.3969/j.issn.1002-4956.2010.02.011.
[30]余晶梅.荧光探针法和疏水相互作用层析法分析蛋白表面疏水性[D].杭州:浙江大学, 2014:28-37.
[31]王辰,江连洲,魏冬旭,等.不同品种大豆分离蛋白结构与表面疏水性的关系[J].食品科学, 2012, 33(9):54-57.
[2]王楚文,柳云龙,唐文静,等.生物保鲜剂对鱼类保鲜的应用现状及其展望[J].安徽农业科学, 2015, 43(16):387-389. DOI:10.13989/j.cnki.0517-6611.2015.16.133.
[3]陈义,汪小钢,宛晓春,等.没食子儿茶素没食子酸酯制备新工艺的研究[J].安徽农业大学学报, 2007, 34(3):364-368. DOI:10.3969/j.issn.1672-352X.2007.03.012.
[4]李鹏,陈莉萍,黄琴,等.EGCG对冷藏罗非鱼鱼片的保鲜[J].食品科技, 2014, 39(8):118-122. DOI:10.13684/j.cnki.spkj.2014.08.027.
[5]骆晓波.表没食子儿茶素没食子酸酯纳米脂质体的制备及其对鳕鱼保鲜效果的研究[D].杭州:中国计量学院, 2015:26-32.
[6]李先宽,李赫宇,李帅,等.白藜芦醇研究进展[J].中草药,2016,47(14):2568-2578. DOI:10.7501/j.issn.0253-2670.2016.14.030.
[7]聂磊,叶敏仪,林静慧,等.白藜芦醇和硝普钠对月季切花的保鲜效应[J].广州城市职业学院学报, 2009, 3(1):44-50. DOI:10.3969/j.issn.1674-0408.2009.01.010.
[8]金宏,惠伟,牛瑞雪,等.白藜芦醇对砀山酥梨的保鲜效应[J].淮北师范大学学报(自然科学版), 2014, 35(1):51-55. DOI:10.3969/j.issn.2095-0691.2014.01.013.
[9]LAILJ,CHIUJM,CHIOURYY.Freshpreservationofalfalfasprouts and mushroom slices by soaking with thymol and resveratrolsolutions[J].FoodScience&Nutrition,2017,5(3):776-783.DOI:10.1002/fsn3.458.
[10]李强.草鱼冷藏过程中肌肉蛋白质的组成和理化特性变化研究[D].长沙:长沙理工大学, 2013:11-21.
[11]朱道立,陈佩林,王康乐,等.骨骼肌肌球蛋白I I的研究进展[J].生物学教学,2010,35(11):8-10.DOI:10.3969/j.issn.1004-7549.2010.11.004.
[12]PARKJAEW,LANIERTC.Scanningcalorimetricbehavioroftilapiamyosinandactinduetoprocessingofmuscleandproteinpurification[J].JournalofFoodScience,1989,54(1):49-51.DOI:10.1111/j.1365-2621.1989.tb08564.x.
[13]LOWRYOH,ROSEBROUGHNJ,FARRAL,etal.Proteinmeasurement with the Folin phenol reagent[J]. Journal of BiologicalChemistry, 1951, 193(1):265-275.
[14]汪金林.茶多酚对冷藏养殖大黄鱼品质影响的研究[D].杭州:浙江工商大学, 2013:33-44.
[15]YEJ,FANF,XUX,etal.Interactionsofblackandgreenteapolyphenols with whole milk[J]. Food Research International, 2013,53(1):449-455. DOI:10.1016/j.foodres.2013.05.033.
[16]TAHAM,QUENTALMV,CORREIAI,etal.Extractionandstabilityofbovineserumalbumin(BSA)usingcholinium-basedGood'sbuffersionicliquids[J].ProcessBiochemistry,2015,50(7):1158-1166. DOI:10.1016/j.procbio.2015.03.020.
[17]RU L, LIU Q, XIONG S, et al. Effects of high intensity unltrasoundonstructuralandphysicochemicalpropertiesofmyosinfromsilvercarp[J]. Ultrasonics Sonochemistry, 2017, 37:150-157. DOI:10.1016/j.ultsonch.2016.12.039.
[18]YAN X, LIU B, CHONG B, et al. Interaction of cefpiramide sodium withbovine hemoglobin and effect of the coexistent metal ion on the protein-drugassociation[J].JournalofLuminescence,2013,142:155-162.DOI:10.1016/j.jlumin.2013.04.009.
[19]王婧.多酚类化合物与牛血清蛋白相互作用的研究[D].杭州:浙江工业大学, 2013:5-10.
[20]BEVINGTONPR,ROBINSONDK,BLAIRJM,etal.Datareduction and error analysis for the physical sciences[J]. Computers inPhysics, 1993, 7(4):415-416. DOI:10.1063/1.4823194.
[21]MANDAL P, BARDHAN M, GANGULY T. A detailed spectroscopicstudy on the interaction of Rhodamine 6G with human hemoglobin[J].Journal of Photochemistry and Photobiology B:Biology, 2010, 99(2):78-86. DOI:10.1016/j.jphotobiol.2010.02.009.
[22]LI J, WANG S. Application of molecular modelling and spectroscopicapproaches for investigating the binding of tanshinone IIA to humanserum albumin[J]. The Journal of Chemical Thermodynamics, 2013,58:206-210. DOI:10.1016/j.jct.2012.11.009.
[23]张淼,王道营,张牧焓,等.一磷酸腺苷与肌动球蛋白相互作用的荧光光谱研究[J].食品科学, 2016, 37(17):32-37. DOI:10.7506/spkx1002-6630-201617006.
[24]林海彬,郑琳,林玉琴,等.邻菲啰啉和氨三乙酸钴配合物与牛血清白蛋白相互作用的荧光分析[J].高等学校化学学报, 2013, 34(8):1818-1825. DOI:10.7503/cjcu20130098.
[25]ROSSPD,SUBRAMANIANS.Thermodynamicsofproteinassociation reactions:forces contributing to stability[J]. Biochemistry,1981, 20(11):3096-3102. DOI:10.1021/bi00514a017.
[26]KLAJNERTB,BRYSZEWSKAM.FluorescencestudiesonPAMAMdendrimersinteractionswithbovineserumalbumin[J].Bioelectrochemistry,2002,55(1):33-35.DOI:10.1016/S1567-5394(01)00170-0.
[27]AZIMIO,EMAMIZ,SALARIH,etal.Probingtheinteractionofhumanserumalbuminwithnorfloxacininthepresenceofhigh-frequencyelectromagneticfields:fluorescencespectroscopyandcircular dichroism investigations[J]. Molecules, 2011, 16(12):9792-9818. DOI:10.3390/molecules16129792.
[28]CHAMANI J, TAFRISHI N, MOMEN-HERAVI M. Characterizationoftheinteractionbetweenhumanlactoferrinandlomefloxacinatphysiologicalcondition:multi-spectroscopicandmodelingdescription[J].JournalofLuminescence,2010,130(7):1160-1168.DOI:10.1016/j.jlumin.2010.02.014.
[29]尹燕霞,向本琼,佟丽.荧光光谱法在蛋白质研究中的应用[J].实验技术与管理,2010,27(2):33-36;40.DOI:10.3969/j.issn.1002-4956.2010.02.011.
[30]余晶梅.荧光探针法和疏水相互作用层析法分析蛋白表面疏水性[D].杭州:浙江大学, 2014:28-37.
[31]王辰,江连洲,魏冬旭,等.不同品种大豆分离蛋白结构与表面疏水性的关系[J].食品科学, 2012, 33(9):54-57.