光谱法研究FCLA与人血清白蛋白的相互作用  

The Study on the Inueraction of FCLA with Human Serum Albumin with Spectroscopy Techniques

作　　者：周静[1] 魏言春[1] 

机构地区：[1]华南师范大学激光生命科学研究所,激光生命科学教育部重点实验室,广东广州510631

出　　处：《光谱学与光谱分析》2007年第11期2287-2290,共4页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(60378043;30470494);广东省自然科学基金项目(015012;04010394);广东省科技计划项目(2004B10401011)资助

摘　　要：在生物学和医学中活性氧的检测非常重要。海萤荧光素类似物FCLA是检测单态氧(1O2)和超氧阴离子(O2-)的一种高灵敏的化学发光探针。文章用光谱法研究了FCLA与人血清白蛋白(HSA)的结合反应,发现FCLA对HSA的荧光有很强的猝灭作用。从荧光猝灭结果求得了FCLA与HSA的结合位点数。根据Frster非辐射能量转移机理探讨了二者相互结合时其给体-受体间的距离和能量转移效率,从而证实了它们的结合作用为静态猝灭过程。阐明了由于能量转移所导致的HSA荧光的猝灭及FCLA荧光的产生机理。通过光谱法对FCLA与HSA的相互作用的研究,阐明FCLA的运输过程及其在体内的作用机制,对实现利用探针在体检测活性氧具有一定的意义,同时也为药物动力分析、体内药物转运的研究提供了基础。It is very important to detect reactive oxygen species （ROS） in biologic and medicinal research. FCLA, the Fluoresceinyl Cypridina Luciferin Analog, is a high sensitivity probe of detecting singlet oxygen and superoxide anions. The FCLA molecule can react with ROS and, after a series of reactions that result in the release of additional chemical energy, releases photons. Depending on the probe used, the photons are typically in visible wavelength range and easily measurable with conventional optical detectors. The binding reaction of FCLA with human serum albumin （HSA） was studied with spectroscopy techniques. It was found that HSA fluorescence was partially quenched by FCLA. The binding constants were obtained with quenching results. According to the Forster energy transfer mechanism, the energy efficiency and the distance between FCLA and HSA were calculated. Then it was found that static quenching was the main binding reaction between FCLA and HSA, and the energy transfer caused the HSA fluorescence quenching and the FCLA fluorescence producing. By studying the binding reaction of FCLA with HSA by spectroscopy techniques, this research proved the transfer style of FCLA and the interaction mechanism in the body. The results will offer some good advises about the ROS detection with the chemiluminesence probe in vivo. Meanwhile, it will also provide a new means to study pharmacokinetics and some basic theories about the medicine transfer in the body.

关 键 词：FCLA 人血清白蛋白 荧光猝灭 结合反应 

分 类 号：O657.3[理学—分析化学]