间氨基苯硼酸与牛血清白蛋白相互作用的研究.pdf

1 间氨基苯硼酸与牛血清白蛋白相互作用的研究 王华芳 1，李文友*1，何锡文 1，张玉奎 1, 2 （1．南开大学化学系，天津 300071，中国；2．中国科学院大连化学物理所，大连 116011， 中国） 摘 要 为了了解分子印迹反应的机理和最适宜的反应条件，应用荧光猝灭法研究间氨基苯硼酸（APBA）与 牛血清白蛋白（BSA）的相互作用。其结果表明：适宜的离子强度和 pH 值为 6.25 时，APBA 与 BSA 的色氨酸 残基的荧光猝灭反应的摩尔比为 2∶1，表观结合常数 KA＝1.0×10 11 L 2 ·mol -2 。通过上述研究，有助于分离或 富集蛋白质中 BSA 组分，从而能够改进印迹和洗脱的效率。 关键词 间氨基苯硼酸；牛血清白蛋白；荧光猝灭；分子印迹 Investigations of the Interaction between 3-Aminophenylboronic Acid and Bovine Serum Albumin WANG Hua-fang1 , LI Wen-you* 1 , HE Xi-wen1 , ZHANG Yu-kui 1,2 (1. Department of Chemistry, Nankai University, Tianjin 300071, China; 2. Dalian Institution of Chemical physics, The Chinese Academy of Science, Dalian 116011, China) Abstract In order to understand the chemical mechanism and optimal conditions of the molecular imprinting, the interaction between 3-aminophenylboronic acid and bovine serum albumin was investigated using fluorescence quenching method with ____________________________________ 收稿日期：2006-08-24；国家自然科学基金(Nos.、天津市自然科学基金(No. 06YFJMJC02800)及湖南大学化学 生物传感与计量学国家重点实验室开放基金(No. 2005009)资助项目. 作者简介：李文友，1966年出生，博士，教授，E-mail: wyli@nankai.edu.cn 2 many factors, as the pH value, ionic strength and others, which maybe influence the interaction. The research results indicate that under appropriate ionic strength, the fluorescence quenching shows APBA strongly bound the tryptophan of BSA with the molar ratio of 2:1 at pH6.25. Their apparent binding constant is KA＝1.0×1011 L2·mol-2, so their chemical binds between donor BSA and acceptor APBA are relatively strong. According to the results, it is predicted the interaction mechanism between APBA and BSA. During following research, it will be entirely possible to separate or enrich BSA composition; maybe it could improve imprinting and eluting efficiency greatly. Keywords 3-aminophenylboronic acid (APBA); bovine serum albumin (BSA); fluorescence quenching; molecularly imprinting 世人关注的蛋白质组学(Proteomics)是在细胞整体蛋白质水平上进行研究，从蛋白质整