胃蛋白酶与对磺酸基杯芳烃相互作用的研究A4.docVIP

胃蛋白酶与对磺酸基杯芳烃相互作用的研究 刘睿*1,3，周兵川2，张军伟1，靳朝喜1，林琳3（郑州大学化学系，郑州 4500）A 文章编号： Study on the interaction between 4-Sulfonic calixarene and pepsin. LIU Rui*1,3,ZHOU Bing-chuan2,ZHANG Jun-wei1,JIN Chao-xi1,LIN Lin3, (1.Luoyang Environmental Monitoring Station,Luoyang 471000,China;2.Songxian Environmental Monitoring Station,Luoyang 471000,China;3.Department of Chemistry, Zhengzhou University, Zhengzhou 450052,China) Abstract: Spectrum Properties study on the interaction of 4-Sulfonic calixarene and 4-Sulfonic calix arene with pepsin were investigated by fluorescence spectroscopy and UV-vis absorption spectroscopy. We found that the two 4-Sulfonic calixarene could quench the fluorescence of pepsin. Based on the Stern-Volmer equation and UV-vis spectrum, the quenching mechanisms were all static quenching. The thermodynamic parameters were calculated, and indicated that the electrostatic forces played a major role in the interaction between 4-Sulfonic calixarene and pepsin. The distance r between the donor (pepsin) and the acceptor (4-Sulfonic calixarene) were determined according to F?rster non-radiative energy transfer theory. The effect of 4-Sulfonic calixarene on the conformation of pepsin was investigated by synchronous fluorescence spectrum. Key words: Pepsin; 4-Sulfonic calixarene; Fluorescence spectroscopy; Interaction 胃蛋白酶（Pepsin）是胃液中主要的蛋白酶，属于水解蛋白酶中的羧基蛋白酶，因含有酪氨酸、色氨酸和苯丙氨酸残基而能发射荧光。胃蛋白酶自 Northrop 1929年将其结晶以来[1]在生命科学[2]、食品科学[3]、分析化学[4]等领域得到广泛的研究杯芳烃是一类由苯酚和甲醛缩合而成的大环化合物,由于具有与环糊精和冠醚相似的性质而被誉为第三代主体分子。水溶性杯芳烃于1955年Cornforth发现具有抗结核的性质[5]，从此对于水溶性杯芳烃在制药方面的应用越来越受到研究者的关注[]。 本文在模拟人体生理条件（pH为2.6，离子强度为0.1 mol·L-1）下，采用光谱技术，研究对磺酸基杯[4]芳烃（简称为SCX4）和对磺酸基杯[6]芳烃（简称为SCX6）胃蛋白酶（简称为Pep）的相互作用，计算了猝灭常数和热力学参数，同时考察了两种对磺酸基杯芳烃对Pep构象的影响。为水溶性杯芳烃在及药物在体内的情况提供的实验数据。1 实验部分 1.1仪器与试剂 日立F-3010型荧光光度计；UNICO UV-2102 PC型紫外可见分光光度计；HANA PH 213型度计；TB-85型恒温水浴锅。Pep（中国医药上海化学试剂公司）：配制成1×10-4 mol·L-1的储备液并于4℃冰箱保存；1.00×10-3 mol·L-1SCX4、SCX6溶液（Acros Organics公司）；pH=2.6的 mol·L-1NaAc-HAc缓冲溶液（内含0.1 mol·L-1的NaCl溶液）；所用试剂均为分析纯以上，水为二次