用圆二色性和荧光光谱技术研究纳米吡啰红G核-铁蛋白的构建机理  

Construction Mechanism of Nanometer Pyronine G Core-Ferritin Revealed with Circular Dichroism and Fluorometry

作　　者：陈盈盈[1] 黄琳[1,3] 季学涛[1] 林青[1] 陈平[1] 柯才焕[2] 黄河清[1,2,3] 

机构地区：[1]厦门大学生命科学学院生物化学与生物技术学系 [2]厦门大学海洋与环境科学学院,近海海洋环境科学国家重点实验室 [3]厦门大学化学化工学院,福建省化学生物学重点实验室,厦门361005

出　　处：《高等学校化学学报》2010年第5期896-904,共9页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30870515);国家“九七三”计划项目(批准号:2010CB12640)资助

摘　　要：小批量制备了电泳纯鲨鱼肝铁蛋白（Liver ferritin ofSphyrna zygaena,SZLF）,对SZLF的结构与功能进行了研究.在pH=2.0~10.0条件下,选用圆二色性（Circular dichroism,CD）光谱技术研究了SZLF和脱铁核SZLF（apoSZLF）二级结构转换的基本特征和变化趋势,揭示了SZLF蛋白壳的亚基稳定性、相互作用强度和去折叠现象.利用酸碱中和方法,解离SZLF蛋白壳亚基,并再次构建成完整SZLF.用紫外-可见分光光度法和荧光光度法研究了构建纳米吡啰红G核-铁蛋白的途径.定量分析结果表明,每分子apoSZLF可捕获12分子吡啰红G于蛋白壳内,构建纳米吡啰红G核-铁蛋白.分析了apoSZLF直接捕获和释放吡啰红G的途径与速率,为后续构建高存量纳米顺铂核-铁蛋白药物载体提供了可行性技术.Liver ferritin of Sphyrna zygaena（SZLF） with electrophoresis purity was prepared in bath to study its structure and function in batch.Under the condition of pH=2.0-10.0,circular dichroism and fluorometry were used to study the primary structural characteristics and change trend of secondary structure conversion both SZLF and apoSZLF,respectively,and revealed the stability,the interaction intensity,and the unfold behavior among subunits of SZLF shell.Using the approach of acidic and basic neutralization,the subunits of protein shell in SZLF can be disassociated,and these subunits disassociated can be also recombined into another new complete ferritin again,which sets up a reasonable technology for constructing a nanometer pyronine G core-ferritin（NPGCF）.Both UV-Vis spectrophotometry and fluorometry were used to study the construction pathways of NPGCF,giving quantificationally twelve pyronine G molecules that were directly trapped into its protein shell for constructing NPGCF by each apoSZLF molecule.In addition,these techniques not only analyze both the pathway and the rate for trapping and releasing pyronine G,but also construct another new drug carrier of nanometer cisplatin core with high carrying capacity-ferritin.

关 键 词：铁蛋白 结构转换 纳米吡啰红G核-铁蛋白 光谱分析 

分 类 号：O657[理学—分析化学]