溴酚蓝结合蛋白结构及稳定性的研究

在早期对溴酚蓝结合蛋白（ＢＰＢＢＰ）的研究中，我们确定了其在电泳谱中的位置及分子量。目前进行的实验中，我们从不同患者血清中ＢＰＢＢＰ的存在及正常人的血清中ＢＰＢＢＰ的结构和稳定性等方面对ＢＰＢＢＰ进行了测定。测定的结果：（１）在不同患者血清中都不同程度地存在着ＢＰＢＢＰ，这为寻求基因药物载体提供了新思路。（２）不同ｐＨ下，紫外－可见光连续光谱的结果说明ＢＰＢＢＰ与溴酚蓝发生了化学键的结合，并结合稳定；只有当ＢＰＢＢＰ的二级结构被破坏时，方可同溴酚蓝解离；溴酚蓝结构中的发色团也参与了结合，同时ＢＰＢＢＰ与溴酚蓝的结合需要电荷的存在；（３）荧光光谱和荧光淬灭及温度淬灭的结果：ＢＰＢＢＰ中位于分子外部的Ｔｒｐ，Ｔｙｓ，Ｐｈｅ占Ｔｒｐ，Ｔｙｓ，Ｐｈｅ总量的６０％，内部的占４０％；在７５℃时ＢＰＢＢＰ发生热变性，说明ＢＰＢＢＰ的热稳定性较好，这也是基因药物的应用基础之一。

Research on the Structures and Stability ofBromophenol Blue-Binding Protein

We have detected the band and molecular weight of bromophenol blue binding protein (BPBBP) in the early research.Recent studies in our laboratory revealed that: (1) There were always different levels of amount of BPBBP in different patients′ albumin; (2) At different pH, the result of ultrovoilet visible continuing spectra suggested that the BPBBP binding with bromophenol blue by chemical binding was stable and the BPBBP can apart from bromophenol blue only when the secondary structure of BPBBP was broken; and the colorable mass of bromophenol blue also took part in the binding, meanwhile the bpbbp can not bind with bromophenol blue without electricity; (3) From the result of fluorescence spectrum and fluorescence puenched experiments, we found that there were 60 per cent of Trp, Tys and phe outside the BPBBP and 40 per cent inside; and from the result of temperature puenched experiments we also find that BPBBP denatured at 75 ℃.This result told us that the thermal stability of the BPBBP is more stable and this is one of the bases of application of gene drugs as well.