|
|||||
|
|
| 人源多巴脱羧酶的表达、纯化以及高通量抑制剂筛选模型的建立 | |
| 引用本文： | 张缘缘,邱军强,周越洋,吴方,张庆华. 人源多巴脱羧酶的表达、纯化以及高通量抑制剂筛选模型的建立[J]. 生物学杂志, 2014, 0(6): 6-10 |
| 作者姓名： | 张缘缘 邱军强 周越洋 吴方 张庆华 |
| 作者单位： | 1. 上海海洋大学水产与生命学院省部共建水产种质资源发掘与利用教育部重点实验室，上海201306; 上海交通大学系统生物医学研究院，上海200240 2. 上海海洋大学水产与生命学院省部共建水产种质资源发掘与利用教育部重点实验室,上海,201306 3. 上海交通大学系统生物医学研究院,上海,200240 |
| 基金项目： | 国家自然科学基金青年科学基金(NO.81102377)；上海高校水产学一流学科建设项目；上海市重点学科建设项目资助 |
| 摘 要： | 帕金森病是一种常见的神经系统退行性疾病。多巴脱羧酶（DDC）是帕金森病研究的靶点蛋白之一,但是目前没有高通量的测活模型。因此,需要构建一种高通量多巴脱羧酶抑制剂的筛选模型,用于发现新型抑制剂。采用克隆表达纯化得到多巴脱羧酶和用于酶偶联反应的磷酸烯醇式丙酮酸羧化酶（PEPC）。基于一系列酶联反应将CO2固定,检测其含量,从而测定多巴脱羧酶的活性。结果得到人源多巴脱羧酶和磷酸烯醇式丙酮酸羧化酶的体外纯酶,建立了一种高通量筛选模型,并且从70个天然化合物中,筛选得到2个多巴脱羧酶的抑制剂。成功构建了一种基于体外纯酶高通量多巴脱羧酶抑制剂的筛选模型。 |
| 关 键 词： | 多巴脱羧酶 帕金森病 抑制剂 高通量 筛选模型 |
Expression,purification of human DOPA decarboxylase and the establishment of high-throughput inhibitor screening model |
|
| ZHANG Yuan-yuan,QIU Jun-qiang,ZHOU Yue-yang,WU Fang,ZHANG Qing-hua. Expression,purification of human DOPA decarboxylase and the establishment of high-throughput inhibitor screening model[J]. Journal of Biology, 2014, 0(6): 6-10 | |
| Authors: | ZHANG Yuan-yuan QIU Jun-qiang ZHOU Yue-yang WU Fang ZHANG Qing-hua |
| Affiliation: | ZHANG Yuan-yuan, QIU Jun-qiang, ZHOU Yue-yang , WU Fang, ZHANG Qing-hua( 1. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306; 2. Shanghai Center for System Biomedicine, Shanghai Jiaotong University, Shanghai 200240, China) |
| Abstract: | Parkinson＇s disease is one of common nervous system degenerative diseases. Dopa decarboxylase（ DDC） has been thought as one of the drug target for Parkinson＇s disease,but there is lacking of a high-throughput screening inhibitor model for DDC. To establish a high-throughput screening model for identification novel inhibitors of DDC,cloning,expression and purification of DDC and phosphoenolpyruvate carboxylase（ PEPC）,monitoring the activity of DDC in the coupling reaction was studied. The CO2 decarboxylated by DDC was detected,based on a series of enzyme-linked reactions. Results showed that the pure enzymes of DDC and PEPC were obtained,and 2 inhibitors were found from 70 natural products using the screening model. |
| Keywords: | dopa decarboxylase Parkinson′s disease inhibitors high-throughput screening model |
| 本文献已被 维普 万方数据 等数据库收录！ | |