鹅掌楸碱与牛血清白蛋白的相互作用研究
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摘要
采用荧光猝灭光谱(FS)、圆二色光谱(CD)、红外光谱(IR)研究了鹅掌楸碱(LA)与牛血清白蛋白(BSA)的结合反应。通过研究发现,鹅掌楸碱和BSA的相互作用机制为静态猝灭。计算了不同温度下的结合常数、结合位点及热力学参数,根据热力学参数推断静电作用力是维系复合物稳定的主要作用力;通过探针反应判断结合位点在SiteⅠ处。根据F9ster荧光共振能量转移理论,计算结合距离r=2.58 nm,说明二者之间形成了LA-BSA配合物。通过同步荧光、圆二色及红外光谱证实了鹅掌楸碱的结合导致了BSA的构象发生了改变。红外曲线拟合定量计算结果显示,BSA的α-螺旋百分含量由50.8%降低到46.5%,表明药物的存在明显改变了BSA的二级结构。
The interaction between liriodenine( LA) and bovine serum albumin( BSA) was investigated by fluorescence spectroscopy( FS),circular dichroism spectrum( CD) and IR spectroscopy.The results indicate that the combination reaction of LA with BSA is a static quenching process.The binding constants,the number of binding sites,and corresponding thermodynamic parameters were calculated,respectively.Based on the thermodynamic parameters,it is speculated that the electrostatic forces play a main role in maintenance stability of complex.Binding site is in the site Ⅰ judged by the probe reaction. The binding distance is 2. 58 nm calculated by the foster fluorescence resonance energy transfer theory.The results show that the conformation of BSA changed due to the interaction of LA.The quantitative calculation of infrared curve fitting shows that the percentage of BSA α-helix decreases from 50. 8% to 46. 5%,which suggests that the presence of LA significantly changes the secondary structure of BSA.
The interaction between liriodenine( LA) and bovine serum albumin( BSA) was investigated by fluorescence spectroscopy( FS),circular dichroism spectrum( CD) and IR spectroscopy.The results indicate that the combination reaction of LA with BSA is a static quenching process.The binding constants,the number of binding sites,and corresponding thermodynamic parameters were calculated,respectively.Based on the thermodynamic parameters,it is speculated that the electrostatic forces play a main role in maintenance stability of complex.Binding site is in the site Ⅰ judged by the probe reaction. The binding distance is 2. 58 nm calculated by the foster fluorescence resonance energy transfer theory.The results show that the conformation of BSA changed due to the interaction of LA.The quantitative calculation of infrared curve fitting shows that the percentage of BSA α-helix decreases from 50. 8% to 46. 5%,which suggests that the presence of LA significantly changes the secondary structure of BSA.
引文
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[2]陈振锋,彭艳,谭明雄,等.基于中药活性成分的金属基抗肿瘤药物前期研究[J].化学进展,2009,21(5):929-933.
[3]CHANG H C,CHANG Fang-rong,WU Yang-chang,et al.Anti-cancer effect of liriodenine on human lung cancer cells[J].Kaohs J.Med.Sci.,2004,20(8):365-370.
[4]WOO S H,SUN Nan-jun,CASSADY J M,et al.TopoisomeraseⅡinhibition by aporphine alkaloids[J].Biochem.Pharmacol.,1999,57(10):1 141-1 145.
[5]刘华钢,秦三海,刘丽敏,等.鹅掌楸碱银配合物体外诱导肺癌SPC-A-1细胞凋亡及其机制的研究[J].中国药理学通报,2008,24(11):1 449-1 452.
[6]林钧材.血液生物化学[M].北京:人民卫生出版社,1988.
[7]孔德贤,周学酬,王玉林,等.结晶紫与牛血清蛋白相互作用研究[J].海峡药学,2013,25(7):200-202.
[8]张海蓉.紫外线照射下Co(Ⅱ)、Pb(Ⅱ)及桂皮酸、青藤碱、儿茶素与牛血清白蛋白的相互作用研究[D].广西:广西师范大学,2007.
[9]CHEN Zhen-feng,LIU Yan-cheng,LIU Li-min,et al.Potential new inorganic antitumour agents from combining the anticancer traditional Chinese medicine(TCM)liriodenine with metal ions,and DNA binding studies[J].Dalton Trans.,2008,2(2):262-272.
[10]黄贤智,郑年梓,陈国珍,等.荧光分析法(第2版)[M].北京:科学出版社,1975.
[11]HU Xiao-xi,DENG Zi-wei,WANG Yun,et al.Study on the interaction between bovine serum albumin and terbium complex containing polymer[J].Soft Mater.,2014,12(3):277-283.
[12]冯小强,李小芳,杨声,等.桂皮酸-1,10-二氮杂菲镧配合物的合成及与牛血清白蛋白的相互作用[J].化学学报,2011,69(16):1 946-1 950.
[13]杨树平,韩立军,潘燕,等.N,N'-双(4-氯苄基)-1,2-丙二胺合铜(Ⅱ)配合物的合成、晶体结构、抑菌活性及与BSA的相互作用[J].高等学校化学学报,2012,33(1):14-21.
[14]王迎进,张艳青,李亚雄,等.光谱法研究根皮苷与人血清白蛋白相互作用[J].分析测试学报,2013,32(2):239-243.
[15]张静,向迅,方婷,等.吲哚美辛与牛血清白蛋白相互作用的光谱研究[J].化学研究与应用,2011,23(7):834-839.
[16]徐同宽,沈兴海,高宏成.光谱法研究3H吲哚季铵盐探针分子与牛血清蛋白的相互作用[J].光谱学与光谱分析,2005,25(6):857-860.
[17]易平贵,刘俊峰,商志才,等.荧光光谱法研究亚甲基蓝与蛋白质的结合反应[J].光谱学与光谱分析,2001,21(6):826-828.
[18]WANG Chun,WU Qiu-hua,WANG Zhi,et al.Study of the interaction of carbamazepine with bovine serum albumin by fluorescence quenching method[J].Anal.Sci.,2006,22(3):435-438.
[19]杨曼曼,杨频.荧光染料探针与蛋白质结合的研究[J].科学通报,1997,42(12):1 276-1 279.
[20]张海容,郭祀远,李琳,等.荧光法研究司帕沙星与白蛋白的作用[J].光谱学与光谱分析,2001,21(6):829-832.
[21]黄波,邹国林,杨天鸣.阿霉素与牛血清白蛋白结合作用的研究[J].化学学报,2002,60(10):1 867-1 871.
[22]程姣燕,邓志红,黄小艳,等.6-乙基-4-乙氧羰基-2-(4-氰基苯基)喹啉与BSA相互作用研究[J].分析试验室,2016,(5):515-520.
[23]鲁子贤.圆二色性和旋光色散在分子生物学中的应用[M].北京:科学出版社,1987.
[24]HONORE B,PEDERSEN A O.Conformational changes in human serum albumin studied by fluorescence and absorption spectroscopy.Distance measurements as a function of p H and fatty acids[J].Biochem.J.,1989,258(1):199-204.